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Western Blotting
Protein Quantification Assay Optimization
Selecting the Right Assay Type
Total protein assays measure the total amount of protein in a sample, most commonly by changing the optical density of the sample in proportion to the concentration. By creating a standard curve based off of the changes of optical density in samples of known concentration, it is possible to calculate the protein concentration in any sample.
One of the first issues to consider when conducting a total protein assay is the type of assay to use. A primary factor to consider in making your choice is the compatibility of the assay with the samples. The objective is to select an assay that requires the least manipulation or pre-treatment of the samples, and will give good results for your sample type.
Use this comparison chart to help decide which protein assay is right for you
| Assay kit | Advantages | Disadvantages |
|---|---|---|
| Coomassie Plus Protein Assay Kit | Fast, compatible with reducing agents | Incompatible with detergents, yeilds poor resutls with very low concentrations |
| BCA Protein Assay Kit | Compatible with detergents and denaturing agents, low variability | Incompatible with reducing agents |
| Micro BCA Protein Assay Kit | High sensitivity and precision | Sensitive to copper reductants and chelating agents |
Keywords: Western blotting optimization, total protein analysis, protein quantification assay, BCA, commassie, protein silver
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