Table of Contents
Total Protein Assays
Quick Technical Summaries......................................3Introduction..........................................................6
Selection of the Protein Assay......................................................................6Selection of a Protein Standard....................................................................7Standards for Total Protein Assay................................................................8Sample Preparation ....................................................................................9Protein:protein Variation ............................................................................9Compatible and Incompatible Substances ................................................10Compatible Substances Table ..................................................................11Time Considerations ................................................................................13Calculation of Results ..............................................................................13
BCA™-based Protein Assays....................................14
Chemistry of the BCA™ Protein Assay......................................................14Advantages of the BCA™ Protein Assay....................................................15Disadvantages of the BCA™ Protein Assay................................................16BCA™ Protein Assay..................................................................................17Micro BCA™ Protein Assay......................................................................18BCA™, Reducing Agent Compatible..........................................see page 24
Coomassie Dye-based Protein Assays (Bradford Assays) 19
Chemistry of Coomassie-based Protein Assays ........................................19Advantages of Coomassie-based Protein Assays ......................................19Disadvantages of Coomassie-based Protein Assays ................................20General Characteristics of Coomassie-based Protein Assays....................20Coomassie Plus – The Better Bradford™ Assay ........................................21Coomassie (Bradford) Protein Assay ........................................................22Coomassie Dry Protein Assay Plates ........................................................22
Overcoming Interfering Substances............................23
BCA™, Reducing Agent Compatible..........................................................24Compat-Able™ Protein Assays..................................................................25
Modified Lowry Protein Assay..................................26
Chemistry of the Modified Lowry Protein Assay........................................26Advantages of the Modified Lowry Protein Assay......................................27Disadvantages of the Modified Lowry Protein Assay..................................27General Characteristics of the Modified Lowry Protein Assay....................27Modified Lowry Protein Assay..................................................................28
Amine Detection..................................................29
OPA Fluorescent Protein Assay..................................................................29Fluoraldehyde
o
-Phthaladehyde................................................................30
Specific Protein Assays
Histidine-Tagged Proteins......................................31
HisProbe™ HRP........................................................................................31SuperSignal
®
West Pico HisProbe™ Kit....................................................31
Antibodies..........................................................32
Easy-Titer
®
IgG and IgM Assays................................................................32
Proteases..........................................................34
QuantiCleave™ Protease Assays................................................................34
Glycoproteins......................................................35
Glycoprotein Carbohydrate Estimation Assay............................................35
Phosphoproteins..................................................36
Phosphoprotein Phosphate Estimation Assay............................................36PhosphoProbe™-HRP................................................................................37
Peroxide............................................................38
PeroXOquant™ Quantitative Peroxide Assay..............................................38
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