Western Blot Protein Quantification Assays

western protein assay

Introduction

After protein sample preparation, the next step of western blotting is total protein analysis. Total protein analysis is the process of quantifying the protein concentration. Protein concentration data is critical to getting good results on your blot: equal amounts of protein must be loaded into each well of the gel to avoid saturation issues, ghost bands, and other trouble. The concentration data produced by total protein analysis can also perform as a reliable loading control for semi-quantitative western blotting. Boster provides a range of high-quality protein analysis kits so you can get more accurate protein concentration data and better blots.

Procedure

To quantify the protein using the BCA protein assay kit, the first step is to reconstitute the bovine serum albumin standard samples with 0.9% NaCl or PBS solution to create a solution with a known concentration. After preparing the BSA standard solution, create a gradient dilution using the BSA standard solution and the diluent solution included in the kit. Prepare the BCA working reagent using a 50:1 reagent A:B ratio and add it to the BSA gradient dilution. Incubate at room temperature and measure the absorbance of the samples at 562 nm. Plot the standard curve to determine the protein concentration of your protein sample.

For a detailed protocol for total protein analysis, see our Western blot handbook.

Tips

  1. Select the right protein assay for your application; choose between silver staining reagents, BCA assays, and commassie blue stains.
  2. Avoid inconsistent band strengths across wells, streaked bands, and saturation problems by getting accurate protein concentrations and optimizing loading amounts. See our troubleshooting guides for more information.
SKU AR0145
Product name Coomassie (Bradford) Protein Assay Kit(for 100-1500 μg/ml protein)
Form Solution (liquid)
Applications Western blotting, ELISA, immunofluorescence; protein expression assays, protein profiling and characterization; protein quantitation assays
Availability: Ships next business day
SKU AR0146
Product nameBCA Protein Assay Kit(for 20-2000 μg/ml protein)
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
Availability: Ships next business day
SKU AR1110
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
Availability: Ships next business day
SKU AR0140
Pack size1kit (for 10 assays to stain gels of 5 x 8.5cm)
FormLiquid
ApplicationsFor protein staining on polyacrylamide gels.

Availability: Ships in 5-7 business days

SKU AR0170
Pack size100 mL(for 10 assays to stain gels of 5 x 8.5cm)
FormLiquid
ApplicationsFor protein staining on polyacrylamide gels.
Availability: Ships next business day
SKU AR0171
Pack size 1 kit (for 30 assays to stain the gel of 5 X8.5)
FormLiquid
Application To detect protein in polyacrylamide gels

Availability: Ships in 5-7 business days

SKU AR0146-500
Product name BCA Protein Assay Kit(for 20-2000 μg/ml protein)
Size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays

Availability: Ships in 5-7 business days

SKU AR0146-1L
Product name BCA Protein Assay Kit(for 20-2000 μg/ml protein)
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays

Availability: Ships in 5-7 business days

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