Question about high salt TBST in WB - (Sep/26/2006 )
I used normal salt TBST in my WB before and got a high background. Then i swithed to high salt TBST. But now i got nothing at all on my film, even for the positive control which works perfect before. I have repeated WB with high salt TBST twice. Still got nothing. Is that possible high salt TBST even washes away the specific band?
Thanks a lot for your help.
what do you call high salt?
when I had to wash carefully after IP, I increased salts up to 1M. No more contamination, but the signal is highly decreased. Now I go up to 0.5M, I think that'enough for what I do .
I think it depends also on your antibody, try to reduce salt.