Bradford protein assay kit 1

Protein concentration was estimated using Bradford Protein Assay Kit I (Bio-Rad, USA) with Bovine Serum Albumin (BSA) as the standard. Briefly, the BSA stock (2 mg/ml) was diluted to 1 mg/ml. Two hundred microlitres of Bradford dye reagent was pipetted into each well of a 96-well dish. The diluted BSA stock was added to the wells to give a final concentration within the range of 0.2l–1 mg/ml. One microlitre of sample was then added to the wells and absorbance was read at 595 nm after 5 min incubation at room temperature. A BSA standard curve was generated for estimation of protein concentration of the samples. Samples corresponding to 100 μg/ml of protein were subsequently aliquoted into microcentrifuge tubes for the lipid peroxidation and antioxidant enzyme assays.