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Author Topic: Using FITC as a hapten tag for IHC  (Read 2261 times)

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Offline yorker01

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Using FITC as a hapten tag for IHC
« on: March 17, 2010, 03:17:18 PM »
 ???
I have a fairly odd question.  Would a FITC-tag (hapten) survive paraffin processing and embedding, and still be recognized by an antibody (clearly it would not fluoresce)?  I have been feeding fluorescently-labelled peptides and proteins to mice, and attempting to follow the trafficking of each after feeding.  However, there is a substantial amount of autofluorescence in the gut (particualry in the green channel), and the overall tissue architecture has not been great.  I am limited in the tags I can use because they have to not inhibit the binding of the peptide antigens to MHC class I or II (FITC has been ok); plus I have a fairly large amount of FITC labelled antigen that I would like to use.  So what I would like to try, is feeding the labelled protein/peptide, fixing the tissue in formalin, process and embedd in paraffin and then try staining paraffin sections with the rabbit anti-FITC antibody I have and come back with an HRP-labeled secondary ab and use something like DAB to detect for protein expression.  The only thing is, I do not know if FITC will survive the processing and embedding (I know DNP does, but I'd like to use the reagents I already have if possible).

Using FITC as a hapten tag for IHC
« on: March 17, 2010, 03:17:18 PM »

Offline excalibur

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Re: Using FITC as a hapten tag for IHC
« Reply #1 on: March 17, 2010, 09:48:26 PM »
You could try it. I have done lots of IHC on paraffin sections for GFP with AEC chromagen.
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Re: Using FITC as a hapten tag for IHC
« Reply #1 on: March 17, 2010, 09:48:26 PM »