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practical experience with paper based peptide enzyme immunossay???

practical experience with paper based peptide enzyme immunossay??? - Western Blot Forum

practical experience with paper based peptide enzyme immunossay??? - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 03-04-2009, 08:50 PM
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Question practical experience with paper based peptide enzyme immunossay???



Hello,
is anyone in the forum who knows something about PBP-EIA?

I ve got a cellulose membrane with small peptide-spots and want to test a protein which should bind to it.
The membrane was incubated in a Blockbuffer containing the protein, washed with TBS and also incubated with the blockbuffer containing the antibody that binds specifically to the protein.
the detection method is chemiluminzenz (HRP).
I ve got a grey or black membran and the spots ve been white (without a signal).
I have no idea what i ve done wrong.
Could it be that i have to use another blockbuffer? Or should i add more protein (higher concentration) to the solution? Or is there a necessary handle that should do and i do not kwow it?

I hope there is anyone who can help me.
bye, trypsin
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