Detection Methods for Immunoblots

Mehdi Alimadadi mehdia at interchange.ubc.ca
Sun May 7 12:32:00 EST 2000


Dear Colloquies,

I have a very good and perfect bands in the SDS-PAGE gel.  I get a good
transfer on the nylon membrane (90-95%) judging by transferred standards but
I cannot get any or a good band on the membrane when using indirect
immunodetection and using DAB detection solution (I make it myself).

What should I use for making an optimum DAB solution, please mention the
chemicals, the concentrations, and the incubation time.

Again this is the case for the immunohistochemistry that I cannot get a good
staining on the fixed cells (sperm and cancer cells) after using DAB
solution.

What could probably be the reason?  I should at least have staining for the
positive control !!!!!!!

Please kindly let me know and thanks beforehand for all your answers.

Thanks,
Afsaneh Motamed Khorasani
PhD Student
E-mail: afsanehm at interchange.ubc.ca








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