HELP: Western Blot problem - (May/26/2005 )

Hi,
I am doing westerns for a year, but since two weeks ago i can't repeat any western i did before. The problem is that i can't detect any band.

I have tried to change samples, primary antibody, secondary antibody, change the primary antibody dilution, change the gel-type, change the chemiluminiscence kit. I don't know what else to do to detect any band.

I would like to receive any possible solution

Thanks

hi
i think the first step is to do an other wblot with extracts of the good one. one time, i got problems and wasn't able to pass. I've asked a colleague to do it and it worked revaling the key i've changed...

[quote=Cristina,May 26 2005, 01:12 PM]

Thanks for your answer, but i have tried with extrats that were used in good westerns and also other colleague did westerns without any result.. I am completely bloked

Cristina,
howz the developer?? Is it being serviced...Also if you have a lab mate who is ready to help you (if u are lucky ) do parallel blots using the same samples..see how it works with that person.
keep up the spirits..it HAS to work
p..