problem with stripping membranes of western blott - do you know some efficient stripping buffer (Aug/12/2005 )

Hi,
I'm trying to strippe my membranes wich i devlopp for western blott using vectastain kit. the problem is that stripping these membranes doesn't work. I used a (NaOH, HCL) buffer and (mercapto-ethanol) buffer and i still see the first bands. I Want to know if someone did it before with membranes developped with vectastain (Vector) for western.
TANK YOU for your help and sorry for my english.

Hi fonfon,

I strip my membranes regularly and do not have any problem. I use ECL and ECL+ kit from Amersham. My stripping buffer is 2.5mM Tris, pH 6.7, 2% SDS with freshly added Beta Mercaptoethanol(700uL for 100ml of buffer). Immerse the membrane in the stripping buffer for 30 mins at 50C. Then wash well with buffer for an hour before you block and delevop.

Wish you luck
-AB

Hi,
I'm trying to strippe my membranes wich i devlopp for western blott using vectastain kit. the problem is that stripping these membranes doesn't work. I used a (NaOH, HCL) buffer and (mercapto-ethanol) buffer and i still see the first bands. I Want to know if someone did it before with membranes developped with vectastain (Vector) for western.
TANK YOU for your help and sorry for my english.

hi
i use glycine 0.2M tween 0.05% at 80°/20'.
Discussion was here, dealing about stripping and reprobing membranes.
fred

thank you for your help .
thank you viteran and BALAKRAM.
Fonfon