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HOME: Table of content Selected Protocols |
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Absorbance 280nm |
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PAGE-SDS (Laemmli) Protein extraction from Polyacrylamide Gel Protein extraction from Polyacrylamide Gel Gel Stain |
Proteases |
Analytical Ultracentrifugation Protein-Protein Interaction Light Scattering Circular Dichroism |
Protein Quantitation
Comparision of Different
Protein Determination Methods
Absorbance 280nm
According toProtein Protocols in CD Rom
Bradford
Lowry According toProtein Protocols
in CD Rom
Lowry-Peterson For membrane proteins, diluted
solutions or solutions with interferants
Protein Precipitation
Methods to concentrate or eliminate interferences
before electrophoresis or protein determination
ADVANSTA Preparing Samples for
Western Blot Analysis: Protein Quantification. A short explanation about different quantification methods (pdf)
BIO-RAD Poster with a brief description of protein assays from Bio-Rad, and reagent compatibility (pdf)
BIO-RAD
Bradford
Modified BIO RAD protein Assay (pdf)
BIO-RAD Lowry Modified
DC BIO-RAD (pdf)
(pdf-2)
BIO-RAD Lowry
Modified RC DC BIO-RAD (pdf)
EXPEDEON BradfordUltra™
kit is a quick and ready-to-use Coomassie-binding, colorimetric method
for total protein quantitation in an environment
containing up to 1% detergent (1%
high protein range, 0.1% low protein range). Is an improvement over
classical Bradford formulations that cannot
tolerate detergent contamination of the protein samples. (pdf)
G-Bioscience CB-X™ Protein Assay: uses a protein dye that is an improvement on the Bradford Coomassie dye reagent (pdf)
GENO Non-Interfering Protein Assay GENO Technology
INVITROGEN : Comparision of Different
Protein Determination Methods
INVITROGEN : CBQCA
Protein Quantitation Kit High-sensitivity protein quantitation (including lipoproteins
and small peptides) even
in the presence of lipids, detergents,
reducing agents or membranes .The CBQCA reagent becomes fluorescent
upon reaction with amine groups
on proteins (amine of lysine sidechains),
so amines (e.g., Tris or glycine) and ammonium ions should be avoided..
Detects as little as 100 ng of
protein/ml (pdf)
INVITROGEN :
Nano Orange Range between 10 ng/mL and 10 µg/mL. The protein sample is
added to the diluted NanoOrange reagent,
mixture, heated at 95°C for ten minutes and
fluorescence can be measured as soon as the mixture has cooled to
room temperature. Tolerant to
reducing
agents and nucleic acids, not to detergents. Although unusually high
concentrations of lipids in the sample can interfere with the assay,
this
interference
can be eliminated by acetone precipitation of the protein, followed by
delipidation with diethyl ether (pdf)
INVITROGEN :
Quant-iTTM Protein Assay
Kit. Range between 0.25-5 µg of protein. The assay is performed at
room temperature, and the
signal is stable for 3 hours. Common
contaminants, including salts, solvents, 2-mercaptoethanol, amino
acids and DNA, are well tolerated in this assay.
It is not compatible with detergents (pdf) (pdf-II)
INVITROGEN :
FluoReporter
® Biotin Quantitation Assay Kit for biotinylated proteins.
Sensitive fluorometric assay for
accurately determining the number of
biotin labels on a protein. The assay can detect from
4 to 80 pmol of biotin in a sample. (pdf)
INVITROGEN : RediPlate EZQ Protein Quantitation
Kit. Solid-phase, fluorescence-based protein assay that facilitates
fast quantitation
of protein samples prepared for gel
electrophoresis. The assay can be performed in the presence of
detergents, urea and reducing agents.
Spot 1
µL of protein sample onto the prepared paper, stain and
read the fluorescence. Ideal for determining protein concentrations of
samples
prior to polyacrylamide gel electrophoresis.
Effective range for the assay is 0.02–5 mg/mL, or 0.02–5 μg per
spot (pdf)
INVITROGEN : EZQ Phosphoprotein Quantitation Kit (E33201) provides
a fast and simple assay for phosphoproteins in solution.
No radioactivity or antibodies are
required, and sample analysis can typically be completed within
60 minutes. Compatible with samples
containing detergents, reducing agents
and urea buffers with up to 1% carrier ampholytes. Requires only
1 µL of sample, and up to 96 samples,
including standards, can be assayed
simultaneously. This kit is ideal for analyzing protein kinase or
phosphatase activities, as well as for monitoring
relative phosphoprotein concentrations
during chromatography or after IEF fractionation of protein samples.
(pdf)
INVITROGEN : EZQ Phosphopeptide Quantitation
Kit (E33202) permits accurate quantitation of phosphopeptides in
solution, in the
presence of
standard buffer components. No
radioactivity or antibodies are required, and sample analysis
can typically be completed within 60 minutes.
Requires only
1 µL of sample, and up to 96 samples, including standards, can be
assayed simultaneously. This kit is ideal for analyzing phosphatase
and kinase activities, as well as for monitoring
relative phosphopeptide concentrations before and after analysis
by liquid chromatography, mass
spectrometry or other
separation technique. Detection from 0.2–0.8 picomole to about 400
picomoles, depending on the peptide. (pdf)
MERCK: IR-based Protein Quantitation - Results are Independent of Detergents, Reducing Agents and Analysis Time (non-Colorimetric Assay) (pdf)
OZ Biosciences Bradford Protein Assay
Kit. The improved and optimized 1X Bradford reagent buffer
allows superior linearity of response at
low and
high protein concentrations (Low: 0.5 – 50 µg/mL High:
50 – 1500 µg/mL) and determination of protein amount in the
presence
of
detergent (< 0.1%). (pdf)
THERMO: Protein Assay Technical Handbook (pdf)
and Protein Assay Compatibility Table
THERMO: Absorbance 280nm (pdf)
THERMO: Bicinchoninic
Acid KIT (BCA) PIERCE Detergent-compatible formulation based on bicinchoninic
acid (BCA) for the colorimetric detection
and quantitation of total protein. Working
range: 20–2,000 μg/ml. Color formation depends of: the macromolecular
structure of protein, the number
of peptide bonds and the presence of
four particular amino acids (cysteine, cystine, tryptophan and
tyrosine).
Certain substances are known to interfere
with the BCA Assay including those with reducing potential, chelating
agents, lipids and strong acids or
bases. Interfering substances may be eliminated
by dialysis or gel filtration, or diluting the sample until the
substance no longer interferes, or
precipitating the proteins in the sample
with acetone or trichloroacetic acid (TCA) (pdf)
THERMO: Bicinchoninic
Acid KIT (Micro-BCA) PIERCE (pdf)
THERMO: Bicinchoninic
Acid Method (BCA) PIERCE The protein assay combines the well-known
reduction of Cu+2 by protein to Cu+1 in an
alkaline
medium with the cuprous (Cu+1) ion
detecting property of BCA.Color formation depends of: the macromolecular
structure of protein, the number
of peptide bonds and
the presence of four particular amino acids (cysteine, cystine, tryptophan
and tyrosine). Reducing agents and copper chelators
do interfere with this assay. Interfering substances
may be eliminated by dialysis or gel filtration, or diluting the
sample until the substance no longer
interferes, or precipitating the proteins in
the sample with acetone or trichloroacetic acid (TCA)
(pdf)
THERMO: Bicinchoninic
Acid Compatible Substances (BCA) PIERCE
(pdf)
THERMO: BCA™ Protein Assay Kit−Reducing Agent Compatible.
For quantitation of total protein in samples while minimizing
interference
from disulfide reducing agents. A
compatibility reagent that modifies disulfide reducing
agents is added to the sample before adding the
BCA™ Reagents. This assay is also
compatible with most ionic and non-ionic detergents in the
presence of a disulfide reducing agent.
Very useful for membrane proteins. (pdf)
THERMO: Biuret
PIERCE (pdf)
THERMO: Coomasie
Dye Binding KIT PIERCE A modification of the well-known Bradford Coomassie® Dye-protein
binding colorimetric method
for total protein quantitation. Most ionic
and nonionic detergents interfere with the assay. Interfering substances
may be eliminated by dialysis or gel
filtration, or diluting the sample until the
substance no longer interferes, or precipitating the proteins in
the sample with acetone or trichloroacetic acid
(TCA) (pdf)
THERMO: Coomasie
Dye Binding Method PIERCE (pdf)
THERMO: Glycoprotein
Carbohydrate Estimation KIT PIERCE
(pdf)
THERMO: Interfering
substances PIERCE (pdf)
THERMO: Lowry
Method PIERCE (pdf)
THERMO: Lowry
Modified KIT PIERCE (pdf)
THERMO: o-Phthalaldehyde
(OPA) Fluorescent Assay PIERCE (pdf)
THERMO: Protein
Assay Preparation Reagent Compat-AbleTMPIERCE
(pdf)
THERMO: Protein
Assay Interfering Substances PIERCE
(pdf)
ROCHE ESL Protein Assay
(pdf)
SIGMA Glycoproteomics Selection Guide — Labeling and Detection,
Quantitation (pdf)
Gel Electrophoresis
PAGE-SDS (Laemmli)
Basic-Native:
For Acidic and Neutral proteins (pI <
7.0)
Acidic-Native: For Basic proteins (pI > 7.0)
Tricine:
SDS gel for low MW proteins (<25 kDa)
Tricine–SDS-PAGE by Hermann Schägger NATURE PROTOCOLS | VOL.1 NO.1 | 2006 | 17 (pdf)
QPNC-PAGE by Bernd Kastenholz: Quantitative Preparative Native Continuous
Polyacrylamide Gel Electrophoresis (QPNC-PAGE). A highly
efficient
approach to resolve native and denatured metalloproteins (MW 6 to >
200 kDa) in complex protein mixtures. (pdf) (pdf 2017)
Phosphate affinity SDS-PAGE Phosphate affinity polyacrylamide gel electrophoresis using Acrylamide-pendant Phos-Tag for the analysis of
phosphoprotein
isotypes (pdf-I) (pdf-II)
Separation and detection of large phosphoproteins by using Phos-tag SDS-PAGE.
E. Kinoshita, et al., Nature Protocols, 4
(2009) pg.1513. (pdf)
BIORAD Precast Gel Electrophoresis Guide(pdf)
Life Technologies Protein handbook: Protein separation by electrophoresis. (pdf-Chapter I)
ABCAM: Western blotting, a beginner's guide. Sample preparation. Electrophoresis. Transfer of proteins and staining. (pdf-I). Troubleshooting (pdf-II)
BIORAD Protein Blotting Guide (pdf)
INVITROGEN : iBlot® Western Detection Kits - Reduce the immunodetection assay time from as long as 2 days to less than 30 minutes (pdf)
Life Technologies Protein handbook: Western blotting and detection technology (pdf-Chapter II)
Protein extraction from
Polyacrylamide Gel
GeBAflex-tube
Protein Extraction from Polyacrylamide Gel (pdf)
Proteomics
Sample Preparation
AGILENT
The Agilent Multiple Affinity Removal System
for Proteomics Sample Preparation. Designed to remove greater than 98-99%
of six
interfering high-abundant proteins using Affinity-purified polyclonal
antibodies to: albumin, IgG, IgA, transferrin, haptoglobin,
and antitrypsin from human serum or
three proteins (albumin, IgG, and transferrin) from mouse
serum samples (pdf-I)
(pdf-II)
(pdf-III)
(pdf-IV)
(pdf-V)
AGILENT
The Agilent
Multiple Affinity Removal System. Designed to remove six interfering
high-abundant proteins from Monkey
Plasma for Proteomics Sample Preparation
(pdf)
AGILENT Agilent Human 14 Multiple Affinity Removal System Columns
for the Fractionation of High-Abundant Proteins from Human
Proteomic Samples (pdf)
ADVANSTA Afyon SDS-PAGE sample preparation kit is a fast, efficient way to
concentrate protein samples and remove buffer components that may
interfere
with electrophoresis. In less than 10 minutes, samples are ready for SDS-PAGE
or immunoblotting. The Afyon protocol is easily scaled up
and can be used for
routine preparation of samples for electrophoresis. (pdf-ApplicNote) (pdf-Brochure) (pdf-Short Protocol)
(pdf-User Manual) (pdf-Protein Analysis)
ADVANSTA Preparing Samples for
Western Blot Analysis: Protein Quantification. A short explanation about different quantification methods (pdf)
G-Bioscience Sample Preparation - Lysis - Fractionation - Clean-Up & Concentration - Handbook & Selection Guide (pdf)
GE-HEALTHCARE Protein Sample Preparation Handbook (pdf)
Overview of protein sample
preparation
Sample collection, stabilization,
and protein extraction
Increasing detectability of
targeted proteins
Ensuring compatibility in protein
sample preparation workflows
GE-HEALTHCARE 2-D Protein Extraction Buffer. Six buffers are available, supplied as a dry powder which is rehydrated with
the
provided diluent. The optimal buffer will depend on the nature of your sample,
and 2-D Protein Extraction Buffer Trial Kit allows
you to evaluate each
extraction buffer to find the most suitable buffer for your sample. 2-D Protein
Extraction Buffer can also be used
prior to 1-D electrophoresis, or for rehydrating IPG
strips prior to 2-D electrophoresis. (pdf)
GE-HEALTHCARE Mammalian Protein Extraction Buffer and Yeast Protein Extraction Buffer Kit are mild, detergent-based cell
lysis methods to extract total soluble protein,
eliminating the need for mechanical cell lysis, and delivering high
quality protein lysate. (pdf)
BIORAD
ProteoMiner™ Protein Enrichment Technology is based
on treatment of complex protein samples with a large, highly diverse library
of hexapeptides bound to
chromatographic supports. In theory, each unique hexapeptide binds to a
unique protein sequence. Because the
bead capacity limits binding
capacity, high-abundance proteins quickly saturate their ligands and
excess protein is washed out during the
procedure. In contrast,
low-abundance proteins are concentrated on their specific ligands,
thereby decreasing the dynamic range of proteins
in the sample. ProteoMiner
Protein Enrichment Kit utilizes single elution reagent. ProteoMiner
Sequential Elution Kit utilizes multiple elution
reagents sequentially elute
proteins based on different properties. (pdf-I)
(pdf-II)
BIORAD Strategies for Proteomics Sample Preparation (pdf)
CALBIOCHEM
ProteoExtract™ Removal Kits (for Enhancing Resolution of
Low Abundance Proteins) (pdf)
GEBA ProteoCon Kit Rapid and easy protein concentration
by spin column. ProteoConD kit: use beads that are for concentration
of protein in denatured
condition. ProteoConN kit: use
beads that are for concentration of protein in native condition. (pdf)
GEBA Maxi GeBAflex-tube Gel Extraction and Dialysis Kit
(3 ml) Handbook. Applications: Extraction of proteins,
RNA, DNA or oligonucleotides
(>20 nt) from polyacrylamide, agrose or any gel matrix in any
running buffer; dialysis or buffer exchange of volumes between 0.1-3
ml; and
preparation of protein samples for MALDI-MS. (pdf-I) (pdf-II)
MERCK ProteoExtract® Protein
Precipitation Kit for the precipitation and clean-up of proteins (pdf)
NORGEN
ProteoSpin™ Abundant Serum Protein Depletion
Kit. The kit depletes 70% of albumin, 90% of α-antitrypsin,
and 50% of transferin
and haptoglobin, enabling the
visualization of low abundance proteins. The kit is unique in that it
is based on an ion-exchange mechanism, silicon
carbide (SiC), and not the use of
specific antibodies. As a result, the kit can
be used to deplete serum proteins from a wide variety of samples,
including human and various
animals. (pdf)
NORGEN ProteoSpin™ CBED (Concentration,
Buffer Exchange and Desalting) Kit. The simultaneous removal
of salts while concentrating a dilute
protein solution makes the kit a
convenient method for preparing proteins before running many downstream
applications such as SDS-PAGE
and isoelectric focusing.
Purification is based on spin column chromatography using
Norgen’s proprietary resin as the separation matrix. These
large columns are frequently
used to prepare protein samples for structural analysis where larger
amounts of protein are needed, such as
X-ray crystallography, NMR
spectroscopy and other applications. The ProteoSpin™ CBED Maxi
Kit comes with solutions for concentrating
and desalting both acidic and
basic proteins. (pdf for Micro)
(pdf for Maxi)
NOVAGEN A comprehensive selection of sample
preparation tools for three main areas of protein research: expression
proteomics, functional proteomics,
and structural proteomics
(pdf)
ProteoEnrich™
ATP-Binders™ Kit allows group separation of protein kinases
and other ATP-binding proteins, yielding partially purified cell
extracts enriched in active protein
kinases. (pdf-II)
ProteoExtract®Native Membrane
Protein Extraction Kit is designed for the isolation of membrane
proteins from mammalian cells and tissues.
The extremely mild procedure yields
a solution of integral membrane and membrane-associated proteins in
their non-denatured state. The
straightforward, two-step procedure
does not require ultracentrifugation or incubation at elevated
temperatures. (pdf-II)
ProteoExtract Phosphopeptide
Capture Kit enables isolation of phosphorylated peptides derived from
cleaved or digested protein samples or
kinase reactions designed for
phosphorylation site identification. Combined with
the ProteoExtract All-in-One Trypsin Digestion Kit, this kit
has a
unique magnetic affinity resin to
isolate phosphorylated peptides with a high degree
of specificity. (pdf-II)
BugBuster® Master Mix extracts
active, soluble protein with maximum yield. It combines BugBuster
Protein Extraction Reagent with Benzonase®
Nuclease and rLysozyme™
Solution in one convenient reagent. The all-in-one protein
extraction reagent efficiently lyses bacteria and digests
nucleic acids.
ProteoExtract Abundant Protein
Removal Kits. These kits facilitate highly specific depletion of either
albumin or albumin/IgG from serum, plasma,
or cerebrospinal fluid.
Selective removal of high-abundance protein improves the detection of
low-abundance proteins of interest. (pdf-II)
ProteoExtract®
Protein Precipitation Kit. Efficient concentration and sample clean-up.
Provides efficient concentration of proteins and removal
of interfering substances from
dilute protein samples in a single step. The ProteoExtract Protein
Precipitation Kit uses a unique combination of
precipitation agents to
quantitatively precipitate proteins and remove interfering substances,
for one-step concentration and clean-up of proteins
(pdf-II)
NOVAGEN Preparation of Protein sample for SDS-PAGE
Procedures and Tips - Article (pdf)
Pall Protein Sample Preparation and Analysis Application Manual. Table of contents (pdf)
1.0 Introduction: 1.1 Proteomics Process Flo -. 1.2 Product Selection (pdf1)
2.0 Pre-Analytical Applications: 2.1
Abundant Protein Removal - 2.2 Protein Fractionation - 2.3 Detergent Removal
- 2.4 Concentration, Desalting,
and Buffer
Exchange - 2.5 Generic Clarification of Samples by Microporous
Filtration (Particulate Removal) - 2.6 Endotoxin Removal (pdf2)
3.0 Analysis: 3.1
Western Blotting - 3.2 Affinity Activated or Activatable Membranes (pdf3)
4.0 Purification Applications: 4.1
Mustang® Ion Exchange Membranes - 4.2 BioSepra® Chromatography
Resins - 4.3 Rapid Purification
Development/Scouting (Process Proteomics) - 4.4 Buffer Exchange,
Desalting, and Concentration - 4.5 Final Product Clarification (pdf4)
5.0 Supporting Technologies: 5.1 Water Supply - 5.2
Microbiology Testing (pdf5)
6.0 Technical Appendices: 6.1 Evaluation of
Chromatography Column Packing Efficiency - 6.2 Optional
Pre-Treatment to Improve Recovery
of Samples
from Ultrafiltration Spin Filters
- 6.3 Vacuum Manifold for Use with Multi-Well Filter Plates -
6.4 General Filtration Procedures
for
AcroPrep™ and AcroWell™ Filter Plates -
6.5 Chromatography Products Selection Guide - 6.6 BioSepra® Media -
6.7
Protein
Sample
Preparation and Analysis Media - 6.8 Chemical Compatibility Guide (pdf6)
Pall Enchant™ Albumin Depletion Kit. The albumin depleting discs
utilize a Cibacron Blue based support that is re-hydrated to form a
gel
based slurry. The gel based
slurry is equivalent to 200 μL of resin. The easy five step protocol
allows you to remove 2 mg of albumin from
each sample processed. (pdf)
Pall
Enchant™ IgG Purification and Depletion
Kits (pdf)
Pall Enchant™ Multi-Protein Affinity Separation Kit
includes all of the components necessary to fractionate albumin and
IgG from 50 μL
of normal human serum or plasma under native or
denatured conditions. (pdf)
THERMO: Manual: Sample Preparation for 2-DE and Mass Spectrometry
(pdf)
An
Introduction to Mass Spectrometry. Poppers™ Liquid Cell
Lysis Reagents. Mitochondria Isolation Kits. Halt™ Protease
Inhibitor Cocktails.
Zeba™ Micro Desalt Spin
Columns.
2-D Sample Preparation: 2-D Sample Prep for
Membrane Proteins. 2-D Sample Preparation for Nuclear Proteins.
2-D Sample Preparation for
Soluble/Insoluble Proteins. SwellGel® Blue Albumin Removal Kit.
ProteoSeek™ Albumin/IgG Removal Kits.
2-D Protein Molecular Weight
Marker Mix. SuperSignal® Chemiluminescent Substrates.
Imperial™ Protein Stain. SilverSNAP® Stain
II.
GelCode™ Blue Stain
Reagent.
Mass Spec Sample Prep:
In-Solution Tryptic Digestion and Guanidination Kit. In-Gel Tryptic
Digestion Kit and PepClean™ C-18 Spin Columns.
Phosphopeptide
Isolation Kit.
Cross-Linkers and Mass Spectrometry in Protein
Structure and Protein Interaction Analysis. Deuterated Cross-Linking Reagents.
Cross-linking Reagents for the
Analysis of Protein Interactions by Mass Spectrometry Analysis
THERMO: PAGE prepTM Protein Clean-up
and Enrichment Kit (pdf)
THERMO: 2
D Sample Preparation for Membrane Proteins (pdf)
THERMO: 2
D Sample Preparation for Soluble or Insoluble Proteins (pdf)
THERMO: 2
D Sample Preparation for Nuclear Proteins (pdf)
THERMO: SwellGel®
Blue Albumin Removal Kit (pdf)
THERMO: Subcellular
Protein Fractionation Kit enables stepwise separation and preparation
of cytoplasmic, membrane, nuclear soluble, chromatin-bound
and cytoskeletal protein extracts from mammalian
cultured cells or tissue. The first reagent added to a cell pellet
causes selective cell membrane
permeablization, releasing soluble cytoplasmic contents. The second
reagent dissolves plasma, mitochondria and ER/golgi membranes but does
not solubilize nuclear membranes.
After recovering
the intact nuclei by centrifugation, a third reagent yields the soluble
nuclear extract. A second
nuclear
extraction
with micrococcal nuclease is performed to release chromatin-bound
nuclear proteins. The recovered insoluble pellet is then
extracted with
the final reagent to isolate cytoskeletal proteins. (pdf)
QIAGEN: Qproteome
Albumin/IgG Depletion Handbook. Fast and specific removal
of albumin and IgG from human serum and plasma samples.
Based on
monoclonal antibodies that bind HSA and human IgG with high affinity
and specificity. (pdf)
QIAGEN: Qproteome™
Bacterial Protein Preparation Handbook. For preparation of
soluble proteins from bacterial cell cultures (pdf)
QIAGEN: Qproteome
Cell Compartment Handbook. For fast and easy subcellular
fractionation of intact eukaryotic cells. By sequential addition
of
different extraction buffers to a cell pellet, proteins in the
different cellular compartments can be selectively isolated: cytosolic
proteins
plasma membranes and compartmentalized organelles, such as nuclei,
mitochondria, endoplasmic reticulum (ER), and cytoskeletal
proteins. (pdf)
QIAGEN: Qproteome Glycoprotein
Fractionation Handbook. For the fractionation of glycoproteins
in proteomic samples (pdf)
Qproteome Total Glycoprotein Kit. The Total
Glycoprotein Spin Columns in the
Qproteome Total Glycoprotein Kit contain ConA
and WGA
lectins. They are used for a general enrichment of the total
glycoprotein population from a cell or serum sample.
Qproteome Mannose Glycoprotein Kit. The ConA, GNA, and LCH lectin spin
columns
in the Qproteome Mannose Glycoprotein
Kit are
used for specific enrichment of glycoproteins with mannose-rich glycan
moieties. The three lectins each bind different
subclasses of these moieties.
Qproteome Sialic Glycoprotein Kit. The WGA, SNA, and MAL lectin spin
columns in the Qproteome Sialic Glycoprotein Kit
are used
for specific enrichment of glycoproteins with sialicacid-rich glycan
moieties. The three lectins each bind different
subclasses of these moieties.
Qproteome O-Glycan Glycoprotein Kit. The AIL, and PNA lectin spin
columns in the Qproteome O-Glycan Glycoprotein Kit are
used for
specific enrichment of glycoproteins with a glycan structure of the
type that are found on T-antigens. The two lectins each
bind
different subclasses of these glycoproteins.
QIAGEN: Qproteome
GlycoArrays is a simple, rapid, kit-based method for determining the pattern
and relative abundance of specific
mammalian glycosylation epitopes in a glycosylated protein. The
analysis can be performed on crude samples in growth media,
eliminating the need for time-consuming purification and sample
preparation steps.
The technology consists of arrays of selected lectins,
which
are used to determine the glycosylation features of the analyzed
glycoproteins. (pdf)
QIAGEN: Qproteome
Mammalian Protein Prep Kit provides gentle but efficient detergent-based
lysis of mammalian cells to deliver a total protein
fraction
suitable for any downstream application. (pdf)
QIAGEN: Qproteome™ Mitochondria Isolation Handbook. For purification
of mitochondria from eukaryotic cells. (pdf)
QIAGEN: Qproteome™
Nuclear Protein Handbook. For isolation of nuclear and nucleic-acid
binding proteins from eukaryotic cell lysates
(pdf-I) (pdf-II)
QIAGEN Phosphoprotein
Purification Kit. For purification and analysis of
phosphorylated proteins from eukaryotic cells (pdf-I)
(pdf-II)
QIAGEN: Qproteome™
Plasma Membrane Protein Handbook. For fractionation of plasma membrane
proteins from adherent cell culture
` samples. The
kit use a ligand specific for molecules on the plasma
membrane vesicles that allow their elution under native conditions
after
ligand–vesicle complexes are precipitated using magnetic beads
that bind to the ligand. (pdf)
QIAGEN: Qproteome
Soluble Protein Separation Handbook. For fractionation of protein
samples used in proteomics analysis. Sequential addition
of
increasing amounts of Fractionation Buffer precipitates proteins that
can be separated by centrifugation. (pdf)
SIGMA:
ProteoPrep 20 Plasma Immunodepletion Kit is a complete kit with all
necessary reagents and consumable equipment to deplete 20
high abundance proteins from human plasma or serum. This kit is
designed to specifically remove the 20 proteins from human plasma.
Specifically, 8 mL of plasma may be depleted in preparation for
proteomic analysis, two-dimensional electrophoresis (2DE), or
liquid chromatography (LC). (pdf)
(pdf-II). Protein Depletion Guide (pdf-III)
VIVASCIENCE Removal
of DNA from lysates of E. coli cells and
French bean tissue prior to electrophoresis.
(pdf)
VIVASCIENCE Vivapure® Anti-HSA Kit for Human Albumin Depletion
Highly specific human albumin depletion
with unique antibody fragments (pdf-I)
(pdf-II)
Vivapure Anti-HSA
Affinity Resin (pdf-III)
Universal sample preparation method for proteome analysis (2009) Jacek R Wis´niewski, Alexandre Zougman, Nagarjuna Nagaraj &
Matthias Mann NATURE METHODS VOL.6 NO.5 pg.:359 (pdf)
Describe a method, filter-aided sample preparation (FASP), which
combines the advantages of in-gel and in-solution digestion for mass
spectrometry–based proteomics
ADVANSTA AdvanStain
Scarlet is
a fluorescent stain for gels and blots that allows sensitive and quantitative
visualization of proteins;
imaged on any fluorescence
imaging system, including laser- and CCD-based systems, with a detection limit
of less than 1 ng of protein
per band
or spot. Fully compatible with downstream Western blotting or mass spectrometry (pdf-I) (pdf-Manual)
G-Bioscience Protein StainsHandbook- A Selection Guide For Protein Stains (pdf)
G-Bioscience Glycoprotein Staining Kit With Rapidstain™ for Enhanced Glycoprotein & Non Glycoproteins Staining (pdf)
GE-HEALTHCARE
Deep Purple™ Total
Protein Stain (pdfI) (pdfII)
(pdfIII)
Deep Purple Total Protein Stain is a ultrasensitive
fluorescent protein stain that can be successfully applied
to both 1-D and 2-D
electrophoresis gels. Imaging of the gels can be performed with a
variety of instrumentation. Deep Purple has significant benefits
in
comparison with other traditional protein stains such as Sypro
Ruby and silver staining, including a potential increase in
sensitivity
with
clear and even background. It is particularly suitable for
quantitation and amenable for use with 2-D image analysis
software programs.
GE-HEALTHCARE
Fluorescent Protein
Gel Stain (pdf)
SYPRO™ Orange, Red, Ruby, and Tangerine protein gel
stains are one-step fluorescent stains optimal for rapid
and efficient staining
of
one-dimensional (1-D) protein gels. These stains provide
sensitivity that is equivalent to the silver staining method
for 1-D gels (1-2ng/band)
and
enable protein detection that is not affected by the presence
of nucleic acids and lipopolysaccharides. SYPRO Ruby stain
is also suitable
for
two-dimensional (2-D) protein gel staining
GE-HEALTHCARE
Silver Stain Kit
(pdf)
A reagent kit for the fast, easy, reproducible and
sensitive staining of proteins in non-denaturing gels, denaturing
gels containing SDS
and/or
urea, and isoelectric focusing (IEF) gels. Allows detection
of most proteins down to the nanogram range, which is 100
times
more
sensitive than Coomassie Brilliant Blue.
GE-HEALTHCARE
SYPRO Orange and
Red Protein Stain Gel (pdf)
SYPRO Orange and Red protein gel stains are designed
for fast, simple, sensitive staining of proteins in electrophoretic
gels. Detection
of
1-2ng of protein per minigel band; more sensitive than Coomassie™
Brilliant Blue and as sensitive as silver staining. Staining
complete
in
<1 hour. Can be used with standard 300nm UV transilluminator
or a laser scanner. Are not suitable for staining proteins
on blotting
membrane
or in IEF gels and they show reduced sensitivity when staining
proteins on 2-D gels.
GE-HEALTHCARE
SYPRO Tangerine Protein
Stain Gel (pdf)
Is designed for fast (less than 1hour), simple, sensitive
staining of proteins in electrophoretic gels (4-8ng
protein / band). Proteins stained
without
fixation can be used for zymography (in-gel enzyme activity)
assays and can also be eluted from gels and used for further
analysis
like
mass spectrometry. Allow visualization of proteins before
proceeding with Western blotting.
BIORAD: Colloidal
Gold Protein Stain (pdf)
A stabilized gold solution optimized for rapid and
sensitive identification of proteins bound to nitrocellulose
membranes.
BIORAD: SYPRO Orange
Protein Stain (pdf)
Provides a reliable and reproducible fluorescent alternative
to silver and Coomassie® staining (for SDS, native,
and 2-D applications),
with
sensitive in the range of 1–10 ng per band. Requires only
30-60 minutes of staining in a single step. Will not stain
DNA or RNA
contaminants.
It is also cost effective, because it is reusable.
BIORAD: SYPRO
Ruby Protein Stains Instruction Manual (protein
gel, protein blot and IEF stain) (pdf)
The fluorescent stain, SYPRO Ruby protein stain is
available in three application specific formulations: 1) the
SYPRO Ruby protein gel
stain
for staining proteins in acrylamide gels (1-D, 2-D, isolectric
focusing); 2) the SYPRO Ruby protein blot stain for staining
proteins on
nitrocellulose
or PVDF membranes; 3) the SYPRO Ruby IEF stain for staining proteins in
acrylamide gels separated by isoelectric focusing (IEF).
These
formulations have been optimized specifically for each application
and the stains are not interchangeable. The stain does
not interfere with
subsequent
analysis of proteins by Edman-based sequencing or mass
spectrometry.
CLONTECH: Glycoprotein Westen detection kit (pdf)
EXPEDEON InstantBlue™ is a ready-to-use, proprietary Coomassie®
stain that is specially formulated for ultra-fast, sensitive and safe
detection
of your
proteins. Protein gels can be stained in minutes without the need to
wash, fix or destain. (pdf)
G-BIOSCIENCES Protein
Gel Stain Selection Guide: LabSafe GEL Blue™ - FASTsilver™
- FOCUS FASTsilver™ - RAPIDstain™ -
Reversible Copper
Stain™ - Reversible Zinc Stain™ -
Glycoprotein Staining Kit (pdf)
GEBA See
Band Protein Staining Solution (pdf)
Sensitivity: 38 ng protein per band.Washing uses water
only.
INVITROGEN : Coomasie FluorTM Orange Protein Gel Stain
(pdf)
Fast, simple, sensitive staining of proteins in electrophoretic
gels (around 8 ng of protein per band). Stained proteins
can be visualized using a
standard
300 nm UV transilluminator or a laser-based scanner. Is
not recommended for staining proteins in 2-D, IEF, nondenaturing
gels,
or
blotting membranes. Do not stain nucleic acid or lipopolysaccharide.
INVITROGEN : Oligohistidine Gel Stain (pdf-I)
(pdf-II)
(pdf-III)
A fluorescent dye selective for oligohistidine fusion
proteins that can be used directly in an SDS.polyacrylamide
gel, eliminating the need to blot
the
protein to a membrane. Staining is complete within hours,
and as little as 15 ng of a hexahistidine fusion protein can
be detected. After
documenting
the oligohistidine signal, the gel can be stained for
total protein using SYPRO ® Ruby protein gel stain.
INVITROGEN : Pro-Q Amber for selective identification of transmembrane
proteins in gel (pdf)
The stain preferentially stains proteins containing
two or more transmembrane domains. After visualizing transmembrane
proteins, the total
protein
profile can be detected using the SYPRO Ruby protein gel
stain. Is not recommended for staining proteins in 2-D, IEF
or
nondenaturing
gels and is not suitable for staining proteins on blotting membranes.
As little as 10 ng of bacteriorhodopsin can
be detected.
INVITROGEN : Pro-Q Diamond Phosphoprotein Gel Stain (pdf)
A simple, direct method for specifically staining
phosphoproteins in polyacrylamide gels. This stain can be used
with standard SDS polyacrylamide
gels
or also compatible with mass spectrometry. The stain allows
detection of phosphate groups attached to tyrosine, serine
or threonine residues
and
can be visualized using a variety of scanning instruments.
The sensitivity limit ranges from ~1–16 ng/band depending on
the phosphorylation
state
of the protein.
INVITROGEN : Pro-Q Emerald Glycoprotein Stain Kit
(pdf)
Fast, simple and more sensitive than any other nonradioactive
glycoprotein staining technique (it is possible to
detect as little as 300 pg of
glycoprotein
per band, depending on the degree of glycosylation).
Compatible with SYPRO Ruby protein gel stain for dichromatic
staining.
Stained
proteins can be visualized using either UV illumination
or a laser scanner.
INVITROGEN : Pro-Q Emerald 300 Lipopolysaccharide Stain Kit
(pdf)
For detection of lipopolysaccharides (LPS) in gels,
with a sensitivity at least 50 times that of silver staining
(as little as 2 ng of LPS per gel lane in
just
a few hours). Stains the periodate-oxidized carbohydrates
of LPS with a bright green fluorescence that is easy to visualize
using a simple UV
transilluminator.
Contaminating proteins can be easily visualized on the
same gel by using the orange-red–fluorescent SYPRO Ruby
protein gel stain.
INVITROGEN : SYPRO® Orange and SYPRO® Red Protein Gel Stains (pdf)
Can detect 4–8
ng of protein. Not suitable for staining proteins on blotting membranes
and reduced sensitivity when staining proteins in IEF or 2-D gels.
INVITROGEN : SYPRO Rubin Protein Gel Stain (pdf)
Created especially for the analysis of proteins in
2-D polyacrylamide gels. It provides the same low nanogram
sensitivity as the best silver staining
techniques,
but stains more proteins and has a much broader linear
quantitation range. Staining is compatible with mass spectrometry
or
microsequencing. Can be visualized with UV transilluminators or laser
scanners.
PERKIN ELMER Phos-tag™ Gel and Blot Stains for Detection of Protein Phosphorylation
Phos-tag™ is a novel chelate with unparalleled selectivity
and sensitivity for phosphoproteins. It exhibits
excellent affinity for phosphomonoesters of tyrosine, serine and threonine.
Phos-tag™ can be linked
to a variety of fluorophores, allowing a range of
choices in spectral properties and detection hardware. Sensitivity to
˜ 1 ng phosphoprotein.
(pdf-I)
(pdf-II)
THERMO: Gel Code Blue Stain
Reagent (pdf)
Utilizes the colloidal properties of coomassie G-250
dye for protein staining on polyacrylamide gels. Stains
only protein and allows bands to be
viewed
directly on the gel during the staining process. After staining,
a water equilibration step (Water Wash Enhancement™) further
enhances
staining
sensitivity and yields a clear background.
THERMO: Gel Code Color Silver
Stain Kit (pdf)
An improved formulation of the Silver Stain procedure.
Has been successfully used to stain proteins, DNA, RNA, lipids
and polysaccharides.
Along
with the quantitation of protein it also characterizes protein
via color. This color characterization distinguishes overlapping
proteins, allows
for
increased pattern recognition and visualization, and identifies
minor contaminants in samples.
THERMO: Gel Code E-ZincTM
Reversible Stain Kit (pdf)
THERMO: Gel Code Glycoprotein
Staining kit (pdf)
THERMO: Gel Code Phosphoprotein
Staining Kit (pdf)
THERMO: Gel Code Silver SNAPTMStain
Kit (pdf)
THERMO: GelCode
Stains Technical Review (pdf)
THERMO: Imperial™
Protein Stain is a coomassie R-250 dye-based reagent for
protein staining in polyacrylamide gels. This reagent stains only
protein and
allows bands to be viewed directly in the gel during the staining
process (≤ 3 ng). The easy protocol is flexible to meet demanding
time and sensitivity
requirements and uses a simple water wash to yield a clear background.
(pdf)
THERMO: Insta StainTM
Blue Gel Stain paper (pdf)
THERMO: Mem CodeTM
Reversible Protein Stain Kit (pdf)
THERMO: Krypton™
Glycoprotein Staining Kit enables fast and sensitive fluorescent staining
for the detection of glycoproteins. The stain is sensitive
down to 15 ng of glycoprotein per band, depending
upon the nature and degree of glycosylation on the proteins. (pdf)
THERMO: Krypton™ Infrared Protein Stain enables sensitive fluorescent
visualization of proteins separated by 1-D or 2-D SDS-PAGE.
Sensitivity 0.25 ng using the standard protocol
or down to 1 ng using the rapid 60 minute protocol. The stain has an
excellent dynamic range and
is compatible with subsequent analysis by mass
spectrometry. (pdf)
THERMO: Silver Stain Rescue Reagent. Silver Stain Rescue Reagent
enables removal of undesirable background from unevenly
and overly
developed
silver-stained gels. (pdf)
SIGMA Protein Staining Reagents for electrophoresis -
Protein Dyes and Stains Selection Chart (pdf)
Colorimetric and Fluorescent Methods for detection
of proteins, glycoproteins, lipids and/or lipoproteins,
nucleic acids, phospholipids,
etc,
for all kind of electrophoresis systems.
SIGMA Glycoproteomics Selection Guide — Labeling and Detection,
Quantitation (pdf)
Silver
Stain
CALBIOCHEM
Proteases Product Listing (pdf)
JENA BIOSCIENCE
Protease Detection Kit detects a wide variety of proteases,
including serine proteases, cysteine proteases and acid proteases
in biological samples. The
kit is based on a labeled casein derivative as protease substrate (pdf)
MOLECULAR
PROBES Protease Assay
Kit (pdf)
NOVAGEN
Protease Assay Kit (pdf)
ROCHE
Universal
Protease Substrate (pdf)
CARDIFF UNIVERSITY - MOLECULAR CELL
BIOLOGY - EHRMAN LAB Proteases
of E.Coli
Analytical
Ultracentrifugation
Alliance Protein Laboratories: Sedimentation equilibrium is
an analytical ultracentrifugation (AUC) method for measuring protein
molecular masses in
solution and for studying protein-protein interactions. It is particularly valuable
for:
1. establishing whether the
native state of a protein is a monomer, dimer, trimer, etc.
2. measuring the equilibrium
constant (Kd) for association of proteins which reversibly
self-associate to form oligomers
3. measuring the stoichiometry of
complexes between two or more different proteins or between a protein and
a non-protein ligand
4. measuring the equilibrium
constants for reversible protein-protein and protein-ligand interactions
(Website A.P.L.) (file)
Alliance Protein Laboratories: Sedimentation velocity is an
analytical ultracentrifugation (AUC) method that measures the rate at which
molecules move
in response to centrifugal force generated in a centrifuge. This
sedimentation rate provides information about both the molecular mass and the
shape
of molecules. In some cases this technique can also measure diffusion
coefficients and molecular mass. Sedimentation velocity is particularly
valuable for:
1. verifying whether a sample is
entirely homogeneous in mass and conformation
2. detecting aggregates in
protein samples and quantifying the amount of aggregate
3.comparing the conformations
for samples
4. establishing whether the
native state of a protein or peptide is a monomer, dimer, trimer, etc.
5. determining the overall shape
of non-glycosylated protein and peptide molecules in solution
6. measuring the distribution of
sizes in samples which contain a very broad range of sizes
7. detecting changes in protein
conformation, for example partial unfolding or transitions to "molten
globule" states
8. studying the formation and
stoichiometry of tight complexes between proteins
(Website A.P.L.) (file)
G-BIOSCIENCES Protein Cross-Linkers Handbook & Selection Guide (pdf)
IBA GmbH: With
four different determination systems based on Strep-tag•II and
One- STrEP-tag, they provide optimal solutions for in vivo
protein-protein
interaction analysis. (pdf)
THERMO: Protein Interactions - Manual (pdf)
Mellacheruvu D. et al. (2013) Nature Methods 8:
730-736 The CRAPome: a contaminant repository for affinity
purification–mass spectrometry data
(pdf) CRAPome
Circular dichroism
Alliance Protein Laboratories: Circular dichroism (CD)
spectroscopy measures differences in the absorption of left-handed polarized
light versus right-handed
polarized light which arise due to structural
asymmetry. The absence of regular structure results in zero CD intensity, while
an ordered structure results in a
spectrum which can contain both positive and
negative signals. (Website A.P.L.) (file)
Alliance Protein Laboratories: (Website A.P.L.) (file)
AVACTA Optim 1000 developed to reduce the time and cost of
therapeutic protein pre-formulation studies, stability testing and formulation.
Thermal unfolding and aggregation
curves are
simultaneously acquired for 48 samples run in a single experiment,
enabling 96
samples to
be analysed in one working day.
Sample volumes as low as 1 μl. Simultaneous
optical measurements: 1) Intrinsic fluorescence to monitor
tertiary structure 2)
Static light scattering to detect aggregates. Sample heating and
cooling
to determine: 1) Protein unfolding temperature
(Tm) 2) Protein aggregation onset temperature (Tagg)
3) Time dependent unfolding and aggregation at a
fixed temperature (pdf)
WYATT Solutions Essential techniques
for characterizing macromolecules and nanoparticles in
solution, to determine molar mass, size, charge,
and interactions
SEC-MALS Conventional Size-Exclusion Chromatography (SEC) relies on reference standards that do not
accurately represent your
molecules. SEC + Multi-Angle Static Light Scattering
(SEC-MALS) measures molar mass directly, independent of elution time, so you
can
have confidence in the values you report.
Molar Mass Multi -Angle
static Light Scattering (MALS) measures molar mass directly, in solution.
Combined with a fractionation technique
like SEC or FFF, MALS determines
absolute molar mass distributions - independent of elution time
DLS Dynamic Light Scattering (DLS) is popular wherever the size and size distributions of
macromolecules and nanoparticles need to
be measured quickly and easily / high-throughput
Nanoparticle Size The size of macromolecules or nanoparticles is
measured via two light scattering methods: Multi-Angle static Light
Scattering
(MALS) and Dynamic Light Scattering (DLS).
TREOS The Multi-Angle Light Scattering
Detector for Absolute Macromolecular Analysis. The miniDAWN TREOS Multi-Angle static
Light Scattering (MALS) detector performs
absolute characterization of the molar mass and size of macromolecules and nanoparticles in solution
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