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Brian J Murphy bmurphy at U.WASHINGTON.EDU
Tue Dec 19 19:10:50 EST 1995


> How to elute antigens without eluting Ab off Prot. A?
> 
> Noam Harel (Harel_n at a1.mscf.upenn.edu@in)
> 18 Dec 1995 02:53:13 GMT
> 
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> --------------------------------------------------------------------
> 
> To: methods-and-reagents at net.bio.net
> 
> From: Noam Harel <Harel_n at a1.mscf.upenn.edu@in>
> 
> Subject: How to elute antigens without eluting Ab off Prot. A?
> 
> Date: 18 Dec 1995 02:53:13 GMT
> 
> Hi,
> 
> I am performing immunoprecipitations on unlabeled nuclear
> extracts using non-covalently linked Protein A-antibody
> combinations.
> The problem is, when I run the precipitates on SDS-PAGE, then do a
> Western blot, the secondary antibody on the W. blot causes the heavy
> chains of the Abs used in the ip to create a
> huge smear that covers up many of the potential bands I'd like to
> see.
> Is there any way besides covalently linking the Ab to the beads,
> or besides eluting with competitor peptides, to keep the Ab from
> coming off the Prot. A beads when boiling in SDS-PAGE buffer?
> I heard something about elution using glycine, but can't find a
> protocol
> Thanks for anyhelp,
> 
> N. Harel
> --------------------------------------------------------------------
> 
>    *  Next message: Patrick HJ Falckh: "Re: RNA extraction from skin
>      samples"
>    *  Previous message: Dima Klenchin: "Re: Using Photoshop for
>      densitometry?"
>    *  Next in thread: brett: "Re: How to elute antigens without
>      eluting Ab off Prot. A?"
>    *  Reply: brett: "Re: How to elute antigens without eluting Ab
>      off Prot. A?"






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