We use this recipe to make 1 L of a mild stripping buffer for re-probing western blots:

Glycine 15 g

SDS 1g

Tween20 10 ml

pH to 2.2 and make up to 1 L with ddH20.

Hope that helps.

Thank you elena..

There are many ways to strip a blot. I use the ECL protocol to strip blots, and I have had very good success.

For 100 ml of stripping buffer:

62.5 mM Tris-HCl, pH 6.8

2% SDS

100 mM Beta-Mercaptoethanol

I make this from diluting stock solutions, then adjust the volume up to 100 ml with dH20.

Next you incubate the membrane in the solution at 50C, not shaking, but agitated occasionally for 30 minutes. Then wash two times for 10 minutes with a high volume of TBS-T (or PBS-T) at room temperature. Then you start from your blocking step (as if you just transferred the blot), and re-probe with primary.,,etc,,,