RIPA Buffer turns Bradford into Blue
Posted 01 March 2011 - 04:08 AM
I am using RIPA protein extraction protocol and in my first trial RIPA buffer in the blank of Bradford was blue, I got negative values with this blank. Later I tried each ingredient of RIPA buffer (SDS, NaCl, Tris, DOC, NP40, ddH2O)in Bradford one by one, only SDS and NP40 (2ul) turn Bradford into blue. I freshly used these 2 and eventhough SDS and NP40 turned Bradford by themselves into blue, buffer was ok when I used it in blank. So problem was neither SDS nor NP40 I thought but after a week I have faced with the same problem. Do I need prepare everything fresh each time. Any idea?
Posted 01 March 2011 - 04:30 AM
I used to use Pierce's BCA kit which is optimised for higher concentration of detergents (see link)
Hope this helps
Posted 01 March 2011 - 04:54 AM