Table of content
Comparision of Different Protein Determination Methods
Absorbance 280nm According toProtein Protocols in CD Rom
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
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)
PIERCE Protein Assay Technical Handbook (pdf) and Protein Assay Compatibility Table
PIERCE Absorbance 280nm (pdf)
PIERCE 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)
PIERCE Bicinchoninic Acid KIT (Micro-BCA) PIERCE (pdf)
PIERCE 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)
PIERCE Bicinchoninic Acid Compatible Substances (BCA) PIERCE (pdf)
PIERCE 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)
PIERCE Biuret PIERCE (pdf)
PIERCE 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
PIERCE Coomasie Dye Binding Method PIERCE (pdf)
PIERCE Glycoprotein Carbohydrate Estimation KIT PIERCE (pdf)
PIERCE Interfering substances PIERCE (pdf)
PIERCE Lowry Method PIERCE (pdf)
PIERCE Lowry Modified KIT PIERCE (pdf)
PIERCE o-Phthalaldehyde (OPA) Fluorescent Assay PIERCE (pdf)
PIERCE Protein Assay Preparation Reagent Compat-AbleTMPIERCE (pdf)
PIERCE Protein Assay Interfering Substances PIERCE (pdf)
ROCHE ESL Protein Assay (pdf)
SIGMA Glycoproteomics Selection Guide — Labeling and Detection, Quantitation (pdf)
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)
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
Protein Extraction from Polyacrylamide Gel (pdf)
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
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
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)
PIERCE: 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
PIERCE: PAGE prepTM Protein Clean-up and Enrichment Kit (pdf)
PIERCE: 2 D Sample Preparation for Membrane Proteins (pdf)
PIERCE: 2 D Sample Preparation for Soluble or Insoluble Proteins (pdf)
PIERCE: 2 D Sample Preparation for Nuclear Proteins (pdf)
PIERCE: SwellGel® Blue Albumin Removal Kit (pdf)
PIERCE 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
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
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
Protein Precipitation Methods
to concentrate or eliminate interferences before
electrophoresis or protein determination
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.
PIERCE: 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.
PIERCE: 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.
PIERCE: Gel Code E-ZincTM Reversible Stain Kit (pdf)
PIERCE: Gel Code Glycoprotein Staining kit (pdf)
PIERCE: Gel Code Phosphoprotein Staining Kit (pdf)
PIERCE: Gel Code Silver SNAPTMStain Kit (pdf)
PIERCE: GelCode Stains Technical Review (pdf)
PIERCE: 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)
PIERCE: Insta StainTM Blue Gel Stain paper (pdf)
PIERCE: Mem CodeTM Reversible Protein Stain Kit (pdf)
PIERCE: 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)
PIERCE: 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)
PIERCE: 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)
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
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)
PIERCE: 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
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,
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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