Sathi- When you get the anti-FITC antibody, instead of having it PE conjugated, you could conjugate it to Alexa488, which has a silimar emission to FITC. This will amplify your signal without having to use up another channel. cheers- Regina Regina H. Harley, M.S. Lab Supervisor - Flow Cytometry Facility Center for Vaccine Development University of Maryland - Baltimore 685 West Baltimore Street Baltimore, MD 21201 (410) 706-7376 FAX: (410) 706-6205 rharley@medicine.umaryland.edu >>> Sathiyaseelan <t.sathi@VERIZON.NET> 05/03/02 12:34AM >>> Hi I have a FITC-conjugated peptide that binds to the surface of a cell line and gives only a moderate intensity of FL1 signal (in 2nd decade) when analyzed by flow cytometry. Unfortunately, I do not have a biotin-labelled peptide in order to use a PE-strepavidin second reagent to amplify the signal. Could any one tell me if there is a possibility to use an anti-FITC antibody conjugated to PE as a second reagent to amplify the fluorescent signal. How much signal amplification I could possibly achieve using this PE conjugated-anti-FITC antibody method and can I use another FITC-labeled antibody to label another surface antigen simultaneously? Does anyone know a source of this antibody? I would appreciate all useful info and help on this. Thanks SATHI
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