BCA Protein Assay
Shaun D. Black
SHAUN%JASON.DECNET at relay.the.net
Wed Mar 2 12:29:27 EST 1994
Take a look at Christia Stoscheck's article in Methods Enzymol 1=
50-68 (1990). It contains a wealth of practical information on UV, C=
Coomassie, BCA, and colloidal Gold protein assays. It is easy to fol=
her protocols for each assay. The BCA assay begins on p.60, as follo=
Reagent A: 1g Sodium bicinchoninate (BCA)
2g Sodium Carbonate
0.16g Sodium tartrate
0.4g Sodium hydroxide
Bring to 100 ml volume with water
Adjust pH to 11.25 with 10 M NaOH
Reagent B: 0.4g Copper sulfate (pentahydrate)
Bring to 10 ml with water
Standard Working Reagent (SWR): Mix 100 vol of Reagent A with 2 =
SWR should be stable for 1 week and should be green.
Combine 1ml SWR and 20 uL of sample; mix.
Incubate 30 min at 60=B0C.
Read Absorbance at 562nm; readings stable for about 1 hr.
There is also a recipe given for a BCA micromethod, on p. 61. Hope t=
helps. Cheers, Shaun
== Shaun D. Black, PhD | Internet: shaun%jason.decnet at relay.th=
== Dept. of Biochemistry | University of Texas Health Center, at =
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