This won't solve the problem. After alcohol fixation the soluble GFP leaves the cell - it won't be there to stain with the Ab. You need to get one of the membrane GFP fusion protein clones and use those. They work great. Both have been published in Cytometry - check Andy Beavis as one of the authors. LArry At 03:23 PM 3/14/00 -0400, you wrote: > >I have never used the site before but it was recommended that I try. I am >interested in doing flow cytometry with a anti-GFP antibody. We are using >a method of intracellular staining which requires methanol >permeabilization. this destroys the innate fluorescence of GFP but we >would still like to identify the GFP transfected cells. Has anyone used a >anti-GFP antibody in flow to address this issue. Thanks. > >Martin > >Martin Carroll, M.D. >Hematology-Oncology Division >University of Pennsylvania >BRB II/III, Rm. 708 >421 Curie Blvd. >Philadelphia, PA 19104 > >Phone: 215-573-5217 >Lab: 215-573-7617 >FAX: 215-573-7049 > Larry W. Arnold, Ph.D. Res. Assoc. Prof. Director, Flow Cytometry Facility Department of Microbiology and Immunology CB# 7290 University of North Carolina Chapel Hill, NC 27599 Phone: 919-966-1530 FAX: 919-962-8103
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