Western problems

[creeves]

I do lots of Western blots with HRP-conjugated secondary Abs and chemiluminescent detection. Most typically, I use RIPA to lyse cells or tissues, Invitrogen precast gels (4-12% BT), and Invitrogen PVDF membranes. Usually the results are great, but sometimes I get crappy data, even though I try to do everything the same way as when it worked well. This sort of thing puts me in a really bad mood, because it takes considerable time to do a Western blot experiment. Who knows where I can find a really comprehensive troubleshooting guide for chemiluminescence Western blotting? Of course the primary antibody is important, but I can deal with this issue using the proper controls.

[admin]

Hello Creeves,
what sort of problems do you get?

We have a small troubleshooting section here for western blot on the western blot page,
and there is a bit more at Western Blot

However, if you can post pictures or describe your problem there are many here that can help.

hope we can get you into a better mood!