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?

-Roman80-

I have used this. Works very well for reprobing.

http://www.piercenet.com/products/browse.cfm?fldID=02040210

-udbiochem-

I always used low pH buffer (pH2.4 - 0.1M glycine, If I remember correctly). Haven't done it recently since switching jobs.

-rosewater-

QUOTE (Roman80 @ Jul 29 2008, 07:46 AM)

only NaOH in solution is no buffer; I think conditions too harsh for Western blots;

we successfully use Pierce restore stripping solution

-The Bearer-

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.

-Michelle4-

QUOTE (Michelle4 @ Jul 30 2008, 02:02 AM)

okay, it´s plausible, however, for using a different species specific secondary antibody, you do not have to strip or block again...

-The Bearer-