|From:||Abizar Lakdawalla |
If the protein is not expected to be exported out of the cell, I would try in situ hybridization as a parallel adjunct to the IHC. I do not know how feasible that would be but it would provide very definitive proof.
> -----Original Message-----
> From: Patterson, Noelle (NIDDK) [mailto:NoelleP@intra.niddk.nih.gov]
> Sent: Tuesday, April 02, 2002 9:11 AM
> To: firstname.lastname@example.org
> Subject: Antibody Characterization
> Anyone out there been tasked to characterize the specificity
> of a novel
> antibody using IHC as "proof"?
> If so, what approaches did you use? How did discern "real" from
> background? Particularly since I am to characterize an antibody to a
> protein that is not (at all) understood; and has no
> "expected" pattern of
> expression. I thought about all the standard background blocking
> techniques, but feel I need to get more "sophisticated" to convince
> I welcome any helpful journal references. I have a few, but am always
> looking for "better ways" to do things.
> Noelle Patterson
> Bethesda, MD 20889-5603