Thermo Scientific Pierce Protein Assay Technical Handbook Version 2
Thermo Scientific Pierce Protein Assay Technical Handbook Version 2
Thermo Scientific Pierce Protein Assay Technical Handbook Version 2
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Table of Contents<br />
Total <strong>Protein</strong> <strong>Assay</strong>s<br />
Quick <strong>Technical</strong> Summaries 1<br />
Introduction 4<br />
Selection of the <strong>Protein</strong> <strong>Assay</strong> 4<br />
Selection of a <strong>Protein</strong> Standard 5<br />
Standard Preparation 6<br />
Standards for Total <strong>Protein</strong> <strong>Assay</strong> 7<br />
Compatible and Incompatible Substances 9<br />
Compatible Substances Table 10<br />
Time Considerations 12<br />
Calculation of Results 12<br />
<strong>Thermo</strong> <strong>Scientific</strong> <strong>Pierce</strong> 660nm <strong>Protein</strong> <strong>Assay</strong> 13<br />
Overview 13<br />
Highlights 13<br />
Typical Response Curves 14<br />
BCA-based <strong>Protein</strong> <strong>Assay</strong>s 15<br />
Chemistry of the BCA <strong>Protein</strong> <strong>Assay</strong> 15<br />
Advantages of the BCA <strong>Protein</strong> <strong>Assay</strong> 16<br />
Disadvantages of the BCA <strong>Protein</strong> <strong>Assay</strong> 17<br />
BCA <strong>Protein</strong> <strong>Assay</strong> – Reducing Agent Compatible 18<br />
BCA <strong>Protein</strong> <strong>Assay</strong> 19<br />
Micro BCA <strong>Protein</strong> <strong>Assay</strong> 20<br />
Coomassie Dye-based <strong>Protein</strong> <strong>Assay</strong>s<br />
(Bradford <strong>Assay</strong>s) 21<br />
Chemistry of Coomassie-based <strong>Protein</strong> <strong>Assay</strong>s 21<br />
Advantages of Coomassie-based <strong>Protein</strong> <strong>Assay</strong>s 21<br />
Disadvantages of Coomassie-based <strong>Protein</strong> <strong>Assay</strong>s 21<br />
General Characteristics of Coomassie-based<br />
<strong>Protein</strong> <strong>Assay</strong>s 22<br />
Coomassie Plus (Bradford) <strong>Protein</strong> <strong>Assay</strong> 23<br />
Coomassie (Bradford) <strong>Protein</strong> <strong>Assay</strong> 24<br />
Removal of Interfering Substances 25<br />
<strong>Thermo</strong> <strong>Scientific</strong> Compat-Able <strong>Protein</strong> <strong>Assay</strong>s 26<br />
Modified Lowry <strong>Protein</strong> <strong>Assay</strong> 27<br />
Chemistry of the Modified Lowry <strong>Protein</strong> <strong>Assay</strong> 27<br />
Advantages of the Modified Lowry <strong>Protein</strong> <strong>Assay</strong> 28<br />
Disadvantages of the Modified Lowry <strong>Protein</strong> <strong>Assay</strong> 28<br />
General Characteristics of the Modified Lowry<br />
<strong>Protein</strong> <strong>Assay</strong> 28<br />
Modified Lowry <strong>Protein</strong> <strong>Assay</strong> Reagent 29<br />
Amine Detection 30<br />
OPA Fluorescent <strong>Protein</strong> <strong>Assay</strong> 30<br />
Fluoraldehyde o-Phthaladehyde 31<br />
Specialty <strong>Assay</strong>s<br />
Histidine-tagged <strong>Protein</strong>s 32<br />
<strong>Thermo</strong> <strong>Scientific</strong> HisProbe-HRP Kits 32<br />
Antibodies 33<br />
IgG and IgM <strong>Assay</strong>s 33<br />
Proteases 35<br />
Protease <strong>Assay</strong>s 35<br />
Glycoproteins 36<br />
Glycoprotein Carbohydrate Estimation <strong>Assay</strong> 36<br />
Phosphoproteins 37<br />
Phosphoprotein Phosphate Estimation <strong>Assay</strong> 37<br />
Peroxides 38<br />
Quantitative Peroxide <strong>Assay</strong> 38<br />
Spectrophotometers 39<br />
BioMate 3S UV-Visible Spectrophotometer 39<br />
Evolution 260 Bio UV-Visible Spectrophotometer 39<br />
Evolution 300 UV-Visible Spectrophotometer 40<br />
Evolution Array UV-Visible Spectrophotometer 40
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