chemiluminescence detection/western blot

Dan Oerther doerther at
Mon Feb 13 09:52:54 EST 1995

In article <D3r575.D3u at>, janapati at wrote:

> I am getting high background and multiple proteins are getting
labelled.  I am using monoclonal primary antibodies.  Any suggestions on
blocking agent or detection method.  

   I am using the Boehringer Manheim HRP kit quite easily with PVDF
membrane blocking for one hour with 1% milk (weight/vol) and 0.1% Tween -
20 (vol/vol) in Phosphate Buffered Saline (PBS).  Additionally, I carry
out the hybridizations in the same solutions.  Hope this helps.  By the
way, I also tried Zeta Probe Nitrocellulose membrane - doesn't block as
well, but seems to work okay if you increase to 5% milk (weight/vol).



Dan Oerther
Departments of Biochemistry and Civil Engineering
Northwestern University, Evanston, IL

doerther at

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