a protocol we have had a lot of luck with over the years is coupling the
antibody to Tosyl-activated Dynabeads according to the manufacturers
instructions. In our hands Dynabeads have given minimum non-specific
binding in a broad range of crude lysates from yeast, E. coli, brain, liver
and muscle. They are expensive and we have looked at a number of
alternatives to save our scarce funds but keep coming back to them in spite
of the cost.
I can send you a detailed protocol if you like?
Good luck....Ken
>I am looking for some general protocols for immunoprecipitation or better for
>things never to do or things which always work. It seems that during
>immunoprecipitation, in particular from crude mixtures, unspecificity can be
>rather high and at the same time, yields can be rather low. Are there any
>tricks to help reverting this relationship?
>
>Any input is appreciated.
>
>Kristine
>
>Kristine Swiderek
>Assoc. Research Director, Biological Structure
>ZymoGenetics, Inc.
>1201 Eastlake Ave. E
>Seattle, WA 98102
>Phone: (206)515-4901
>Fax: (206)442-6608
>e-mail: swiderek@zgi.com
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Ken I. Mitchelhill
The John Holt Protein Structure Laboratory
St. Vincent's Institute of Medical Research
41 Victoria Parade
Fitzroy 3065 Victoria
AUSTRALIA
Telephone: 61-3-9288 2480
Facsimile: 61-3-9416 2676
Email: k.mitchelhill@medicine.unimelb.edu.au
Laboratory: http://www.medstv.unimelb.edu.au/WWWDOCS/SVIMRdocs/JHPSL.html
ABRF: http://www.abrf.org
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