Frank Bottone

8/5/2002

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.