restore dried PVDF - (Jan/15/2008 )

Hi,

I used to dry my PVDF for longer storage. I've moved to other lab and the restoration doesn't work

(got all blurry, strong background and no band)

I dip the blot in MeOH for minutes and transfer to TBST, start with blocking.

Does anyone know how to restore dried blot?

-anna_L-

you only need to dip it in the methanol for 20-30 seconds then buffer and continue.

how does the water and other reagent quality compare with that of your previous lab?

-mdfenko-

Thank you. I've worried that I may dip a blot in MeOH too long and wash out all ptn. I'll try again.

Maybe I'd prepare all the reagent and buffers freshly by myself and order new antibody.

Somehow none of lab western did work for months.

-anna_L-

may be the PVDF is old.

-Minnie Mouse-

QUOTE (anna_L @ Jan 15 2008, 05:30 PM)

Hi,

I used to dry my PVDF for longer storage. I've moved to other lab and the restoration doesn't work

(got all blurry, strong background and no band)

I dip the blot in MeOH for minutes and transfer to TBST, start with blocking.

Does anyone know how to restore dried blot?

to re-use once activated PVDF (by methanol) you should not do it a second time, especially if PVDF carries bound proteins...

-The Bearer-