immuno blot

David E Bell bell at mari.acc-admin.stolaf.edu
Wed Feb 10 10:16:15 EST 1993



a simple experiment, but not working...
who can help ???

I did immunoblots (running ag on a gel, blotted it, incubated blot with
		   positive human serum, developed with anti human POD
		   conjugate  ==> no bands !!)
Ponceau staining revealed lots of protein on my blot.
My substrate reaction is positive, too.
After several investigations the problem is bordered in:

	no binding of the antibodies (or not enough) in the positive
	patient serum to the on NC membrane presented antigens !!

why ?? did anybody experience  similar problems ??

in addition it must be said, that i did the same procedure as ELISA (same
antigens coated, same human serum, same POD conjugate !!) - and there it was
working very well !!!

So any ideas what prevents ab binding to NC presented antigens ????

I appreciate every hint, thougt or idea. Thanks.

Tom Pasternak
Immunology University Aberdeen
Internet: ugn4501 at aberdeen.ac.uk

or as sender: bell at stolaf.edu



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