I am a doctoral student at the Faculty of Sciences Rabat Morocco, I am working on the flow cytometry analysis of an intracellular protein on different lymphocytes,
For this purpose I make a surface marking of TCD4 lymphocytes in two different tubes and after cell then I make an intracellular permialization: in one of the tubes I use the isotype of the intracellular protein as control and its conjugate, and on a second tube I use the protein that I am looking for and its conjugate, however as a result I do not get any difference between the protein and its isotype, or as today I obtained the magnitude of the protein larger than that of the isotype, and returning to the literature they always find the histogram of the isotype to a greater extent than that of the protein? please, what do you think about that result ?
-I have a second question about the kaluza analysis software : could you tell me how to replace two different graph (Isotype and poteine) in one FCS file all knowing that each of the two graph is in different FSC file,
Thank you so much