[Histonet] custom-made polyclonal antibody doesn't stain

Barbara Verstraeten Barbara.Verstraeten <@t> ugent.be
Tue Jan 4 08:14:27 CST 2011

Dear all,


I had two antibodies made (in rabbit) for the same peptide.

It was the first time I did this : the firm designed the immunization
peptide, injected and bleed the rabbits and purified the antibody.

I have tested it on paraffin sections: all background, no staining where it
should be. I tried different retriever buffers and detection with dab and

I have tested it on western blot: a lot of background, dirty lanes. To get a
clearer view I performed a co-IP. Still not really great.


Can anybody help me on this issue?

I just want it to work for staining. 


Thanks a lot in advance!


Barbara Verstraeten, Drs.

Evolutionary Developmental Biology

Department of Biology

Ghent University

K.L. Ledeganckstraat 35

B-9000 Ghent


tel: ++32/(0)9 264 52 31

fax: ++32/(0)9 264 53 44



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