Re: Using Human antibodies for IHC on HUMAN tissue
|From:||Abizar Lakdawalla <firstname.lastname@example.org>|
we do have a human on human ihc product (similar to our iso-IHC for
mouse on mouse).
Works pretty well with humanized antibodies on human tissues.
Miriam Schroeder wrote:
> Hi all. I have a request to use a HUMAN antibody as
> the primary antibody for staining both mouse tissue
> (shouldn't be a problem) and HUMAN tissue.
> I am anticipating staining the human tissue will be
> problematic due to 1)Secondary (anti-human) antibody
> reacting with endogenous antibodies already in the
> tissue AND/OR 2)Capture of the primary antibody by Fc
> My usual procedure for IHC is to use a biotinylated
> secondary, then streptavidin-peroxidase conjugate &
> finally DAB. I am thinking if there were a way to
> biotinylate my primary antibody (prior to application
> to the tissue) that I could avoid using the
> biotinylated secondary & get around cross-reactivity
> problems that way.
> I am wondering if anyone has a tried-and-true
> procedure for dealing with this sort of situation.
> (Or, preferrably, knows of a kit for biotinylation of
> the primary HUMAN antibodies.) (I've seen lots of
> kits for biotinylating MOUSE primaries, but haven't
> come across any for HUMAN primaries.)
> I realize there may be other ways around this, like
> "block" endogenous antibodies in the tissue with an
> UNLABELED anti-human secondary at the beginning of the
> procedure, pre-adsorbing the secondary (but I'd like
> to steer clear of human serum, don't know where to get
> it anyway), etc. etc. Time is of the essence so I
> don't have time to work out a procedure on my own that
> might employ these type methods.
> Any help from someone with ACTUAL EXPERIENCE in this
> type situation would be greatly appreciated. We can
> all hypothesize about ways around this (see above
> paragraph) until we are blue in the face but at the
> end of the day any such procedure would have to be
> developed/optimized first, and I don't have time for
> Thanks so much!
> Miriam Schroeder
> Research Associate
> Berlex Biosciences
> Richmond, CA
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