Silver staining problem

Viewing 2 reply threads
  • Author
    Posts
    • #14255
      hericorreia
      Participant

      Hi everyone. I have a problem staining an second dimentional gel of proteins. I get negative spots after stain with silver nitrate. I had done different silver staining protocols and the result is always the same. negative spots and dark baground. I have changed the reagents, wash steps, incubation times with fix solution and silver nitrate, protein concentrations- etc.
      Have you any suggestions to resolve this trouble?
      thank you

    • #102693
      JackBean
      Participant

      didn’t you overstain? Did you try to decrease the last time, before you stop the staining with acetic acid? (sorry, I don’t remember these photographic terms). Do you control, how long you have it in that solution or you just have it there for designed time?

    • #102696
      canalon
      Participant

      I’ve had that in a few occasion on simple gel where I had too much protein (intense coomassie staining), the silver stain gave a blank center with only the side lightly colored. But I never had a negative stain on the whole gel. But I use a Bio Rad kit, rather than home made reagent.
      Sorry not much help here. Maybe you should try Coomassie staining ? that is what is recommended by the lab I plan to use for sequencing my protein if I finally manage to express enough of it.
      Sample protocols available here:
      http://www.sickkids.ca/Research/APTC/Ma … index.html

Viewing 2 reply threads
  • You must be logged in to reply to this topic.