reagents

1. 2% Na2CO3 in 0.1 N NaOH
2. 2% NaK Tartrate
3. 1% CuSO4•5H2O
   Mixture before use:  25.00 ml of 1
                                 0.25 ml of 2
                                 0.25 ml of 3
4.  2N Folin reagent (yellow in color)
5.  Standard: Given BSA 1 mg/ml

 procedure

1. Transfer sample containing up to 40 µg of protein to test tubes and bring volume up to 200 µL with H2O.
2. Add 1 mL of mixture and mix well with the Vortex.
3. Incubate for 10 minutes.at Room Temperature.
4. Add 50 µL of Folin reagent, mix well with Vortex.
5. Incubate for 30 minutes at Room Temperature.
6. Read at 540 nm against an H2O blank.