Pierce Coomasie Kit

Working range with BSA 125-1000ug/ml

• Dilute unknown at least 1:1 with water, can do 1:5 1:10 etc
• Do not dilute standards
• Pipette 50ul each standard or unknown sample into labeled test tube
• Do standards and unknowns in duplicate
• Gently mix contents of reagent by inverting bottle several times
• Add 1.5ml Coomassie Plus Reagent to each tube, vortex
• Let stand at room temp 10 minutes
• Set spectrophotometer at 595nm, measure all samples
• Subtract the (average) 595nm blank value from all other values
• Prepare standard curve, linear regression

One example of Standards

Vial Volume Diluent Volume BSA Final Conc Stock=2000ug/ml

A 0 300ul stock 2000ug/ml

B 125ul 375ul stock 1500ug/ml

C 325ul 325ul stock 1000ug/ml

D 175ul 175ul A 750ug/ml

E 325ul 325ul B 500ug/ml

F 325ul 325ul E 250ug/ml

G 325ul 325ul F 125ug/ml

H 400ul 100ul G 25ug/ml

I 400ul 0 0 Blank

• You can also do a serial dilution 2000, 1000, 500, 250 125 62.5
• Note: Can also add 66.67ul sample plus 2 ml Coomassie reagent