I am having troubles using Ponceau S solution for temporary staining of
western blots.
I am definitely sure that my proteins were transferred to the membrane
(rainbow marker confirms that) and in the satisfactory quantity.
The Ponceau S (Sigma) solution was always prepared fresh according to
Current Protocols in Mol. Biol., what means: 0.5g of Ponceau S dissolved in
1ml of acetic acid and diluted to 100ml with ddH20.
The membrane used were PVDF either from Sigma or Biorad.
I have tried with various incubation times from 5 to 30 minutes. Destining
was under stream of the ddH20.
Directly after semi dry transfer, the membrane was rinsed with PBS
(50mM KP, 100mM NaCl) and immersed into Ponceau S solution.
PLEASE DO SOMEONE HAS IDEA WHAT WAS WRONG. Thanks Zeljko.
