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

[Mark Tarango]

Hi Barabara,

I'd suggest buying an antibody purifcation kit and then try staining with
the antibody.  I've had good luck trying this in the past.

Here is a link to a kit
http://www.piercenet.com/browse.cfm?fldID=4C752E8A-D886-4E12-A2B7-66C00F8B9B2A


Good luck!

Mark

On Tue, Jan 4, 2011 at 6:14 AM, Barbara Verstraeten <
Barbara.Verstraeten <@t> ugent.be> wrote:

> 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
> fluorescence.
>
> 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
>
> Belgium
>
> tel: ++32/(0)9 264 52 31
>
> fax: ++32/(0)9 264 53 44
>
> http://www.evodevo.ugent.be
>
>
>
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