For a 24 well plate:
For more details: https://www.piercenet.com/instructions/2160412.pdf
For a sample Excel spreadsheet, see: BCR WB BCA 2014-8-1.xls
In a 96 well plate, make three rows of protein standards. Add 150uL of protein standard solution and 150uL of working reagent to each well. To make the protein standards:
Vial | Volume of Diluent | Volume and Source of BSA | Final BSA Concentration |
---|---|---|---|
A | 4.5 mL | 0.5 mL of Stock | 200ug/mL |
B | 8.0 mL | 2.0 mL of vial A dilution | 40ug/mL |
C | 4.0 mL | 4.0 mL of vial B dilution | 20ug/mL |
D | 4.0mL | 4.0 mL of vial C dilution | 10ug/mL |
E | 4.0mL | 4.0 mL of vial D dilution | 5ug/mL |
F | 4.0mL | 4.0 mL of vial E dilution | 2.5ug/mL |
G | 4.8mL | 3.2 mL of vial F dilution | 1ug/mL |
H | 4.0mL | 4.0 mL of vial G dilution | 0.5ug/mL |
I | 8.0mL | 0 | 0ug/mL (blank) |
For more details: http://www.bio-rad.com/webroot/web/pdf/lsr/literature/M1703940.pdf
1% NP40
0.1% tween 20
1x HALT protease inhibitor
150 mM NaCl
50 mM Tris
Recommended: make a 10x stock and then dilute to 1x
25 mM Tris, 192 mM glycine, 0.1% SDS, pH 8.3
0.1% SDS means 1g in 1L
48mM Tris, 39mM glycine, (20% methanol) pH 9.2
5.82 g Tris
2.93 g glycine
200 mL methanol
to 1 L with dH2O