How to calculate total protein content (mg/gDWt) after Bradford Assay?

Kiran Reddy @Kiran-Reddy-37

21 March 2025 1 7K Report

Tissue used (dry legume seed powder): 15.7mg, 15.4mg, 15.9mg
BSA concentration for standards (0.5, 1, 1.5, 2 μg/mL)
500μL 0.1M NaOH used as extraction buffer
20μL of protein extract and 200μL Bradford reagent was used

Concentrations (μg/mL) after Absorbance at 595 nm

1. 12.6907

2. 12.6677

3. 12.4587

Satyendra Mondal

Dilute your test samples at least 20-fold so that they are in the range of the standard curve and you can extrapolate the absorbance value in this standard curve. Use more BSA protein concentrations in the lower range, e.g. 0.25, 0.125, 0.025 μg/mL. Also prepare the BSA standard solutions in extraction/lysis buffer.

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