2 replies to this topic

[jasmina]

Hi all,
I tried to strip my filter 2 times with the same stripping solution: 2%sds, 5mM Phosphate buffer, 0.14 per 1000 of mercapto ethanol. incubation 20min at 62°C, then, washes with PBS tween and blocking solution later. when i develop by ECL another another antibody, i could still see old bands on the filter.
i tried to strip another filter using another harsh stripping solution: 2%sds, 62.5mM tris Hcl ph 6.7, 0.8ml mercapto in 100 ml final volume buffer. incubation at 50°C for 45min, then wash with water 1h, then PBS tween, i tried to block and develop ECl to see if there is still signal, so, i still see old bands, may be a little bit lower intense, but still there!
Im wondering if u have idea about better stripping buffer
thanks

[deepgreenbegum]

hi, I used to use Bio-Rad's commercial stripping buffer and it'd work but it'd take a long time (2-3 hours) and needed high temperatures (37-42C). Now I am using Pierce Restore WB Stripping Buffer and it only took 15min at RT to strip everything!

[jasmina]

ok, thank you!
i will inform if they can buy it in the lab