Protein Assay for microplates using BCA kit (Pierce)
Setup standard curve using Pierce #23209 2000ug/ml standard as follows:
ug/ml: use x ul of 2000 ug/ml: up to 1000 ul with dH20:
250 125. 875.
200 100. 900.
150 75. 925.
100 50. 950.
50 25. 975.
25 12.5 987.5
Add 25 ul of each standard to a well in rows A of 96-well dish.
Add 25 ul of dH2O to 2 wells in row A for a blank and a negative control.
Add 23 ul of dH2O to as many wells as you have samples in duplicate.
Add 2 ul of each sample in duplicate to the appropriate wells.
Also do further dilutions if you are unsure if the sample will read within standard curve.
Add 50 ul of BCA solution B to every 200 ul of solution A.
Make enough for a few more samples than you have including the controls and blanks.
Mix gently by hand, incubate for 30-minutes (up to 1 hour) at 37’C. then equilibrate at room temperature before reading at 569nM in a plate reader.