using ABTS with HRP-conj for immunoscreening of lambda library

Bernard Murray bernard at elsie.nci.nih.gov
Sun Jun 26 22:51:44 EST 1994


In article <1994Jun27.004005.9923 at marlin.jcu.edu.au>, robert at chironex.jcu.edu.au writes:
> Hi,
> Just a quick one. Has anyone used ABTS and HRP-conjugated secondary antibodies on Amersham Hybond N+ filters for the purpose of immunoscreening a lambda gt11 
> library ?
> Any pointers would be appreciated.
> Cheers,
> Dr Robert J. Coelen                                  fax:   61-77-791 526
> Dept of Biomedical and Tropical Veterinary Sciences  phone: 61-77-815 024
> James Cook University of North Queensland

Robert,
	Maybe I don't understand the actual protocol you have in mind but
will you not need a substrate that yields a solid precipitate?  ABTS and
other HRP substrates like OPD give soluble products for use in eg. ELISA.
Suitable substrates for use with filters would be diaminobenzidine or
(better) chloronaphthol.  I'm sorry if I'm barking up the wrong tree but
I'm more familiar with using HRP in ELISA and Western immunoblotting.
		Stain on!
			Bernard

Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)



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