Intracellular Assay

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thanks,
gary

We want to detect the synthesis of insulin-like growth factor-I (IGF-I) in
monocytes/macrophages using intracellular flow cytometry. We would like some
advice from colleagues who have experience with this technique or with this
particular application. Ultimately, we would like to sort mixed populations
of differentiating macrophages and characterize the cells that are positive
and negative for IGF-I. For the moment, we would like to get started with a
reliable protocol which works for macrophages, since these cells can have
high background fluorescence. The primary antibody preparation that we have
is the polyclonal rabbit anti-human IGF-I preparation of Underwood which is
distributed by the NIH. One of our concerns is that this antibody is
directed against the mature form of the peptide and may not recognize the
intracellular pre-pro IGF-I.

Any suggestions or advice would be appreciated.

Gary Durack
University of Illinois Biotechnology Center
Voice (217) 244-0559
FAX (217) 244-0466

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CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu