Ponceau S staining problem...

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Thu Nov 30 11:26:50 EST 2000


"Kevin A. Morano" <kevin.a.morano at uth.tmc.edu> wrote:
:
:I find that you need to make a very dark Ponceau stain, on the order of
:a deep cabernet (2-5%??).  It also stains better in 5% acetic acid.  I
:make the stock and reuse it a bunch.  The sensitivity is low, so you
:would need at least 5-10 ug total protein on the blot.
:

5-10 ug? My stock made in TCA/sulfosalicilic acid detects 0.1 ug
without any problem. Surely less sensitive than Coomassie
but good enough so that in most cases I do not need to run
a separate gel for total protein staining. Blot -> stain -> scan ->
destain in pH > 7.5 -> block and do Western.

        - Dima






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