Stripping buffer - (Jul/29/2008 )
Could you tell me please, what is the fastest way to strip the PVDF membrane. Recently, I read about the stripping using NaOH 0.2M solution. And it was mentioned like the stripping takes only 5 minutes. Is it true?
I have used this. Works very well for reprobing.
I always used low pH buffer (pH2.4 - 0.1M glycine, If I remember correctly). Haven't done it recently since switching jobs.
only NaOH in solution is no buffer; I think conditions too harsh for Western blots;
we successfully use Pierce restore stripping solution
0.2M NaOH, 2x 5min. Then wash and block again. This kills the 2ndary antibody but does not actually strip. So it will be ok if you use a difference species antibody to a protein that is different size to the first probing.
okay, it´s plausible, however, for using a different species specific secondary antibody, you do not have to strip or block again...