Detection Methods for Immunoblots

Neal Melvin nrmelvin at ucalgary.ca
Sun May 7 16:03:56 EST 2000


A couple suggestions...

Switch to nitrocellulose membranes instead of nylon...

And use an enhanced chemiluminescence (available from Amersham) detection method
instead of DAB.

Neal

Mehdi Alimadadi wrote:

> 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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