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Quick start Bradford protein Assay (total protein determination)

Updated 7/20/2021 11:39 PM
Step-by-step
by igem_lund

Introduction

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Materials

  • BSA
    • Distilled water
      • BIO-Rad Protein Assay Reagent

      Procedure

      1. Calculate how many samples you will analyze, including one blank. The analysis will beperformed in duplicates.
      1. Prepare a standard series of BSA in the concentrations 1.0, 0.8, 0.6, 0.4, 0.2mg/mL, diluted in DI-water (keep at +4 oC or on ice prior to use).
      1. Prepare a 1X Dye Reagent by diluting the Bio-Rad Protein Assay Dye Reagent Concentrate 5 times with distilled water.
      1. Mix 980 μL of Bradford reagent with 20 μL of blank/standard/sample in a cuvette.
      1. Incubate at room temperature for 5 min.
      1. Measure the absorbance at 595 nm with the provided spectrophotometer.
      1. Subtract absorbance for blank from all other samples.
      1. Create a standard curve for the BSA, and use that to determine the total proteinconcentration in the analyzed protein samples.

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