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Standards (Expression system for standard: E.coli, Immunogen sequence: A2-F222? and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for GSTA is added subsequently and then followed by washing with PBS or TBS buffer Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer HRP substrate TMB was used to visualize HRP enzymatic reaction TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution The density of yellow is proportional to the human GSTA amount of sample captured in plate. Detection Target: Glutathione S-Transferase A1 Uniprot ID: P08263)

Then, careful layering of 2.6 mL each of 1.15 M, 0.86 M, and 0.25 M sucrose solutions was performed on the microsomal suspension, and this step gradient sucrose solution was centrifugated at 82,000 g for 3 h at 4 C