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PerCP/Cy5.5 anti-human CD4 Antibody
317428
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I have used this antibody for analyzing the fraction of CD4+ve T cells in a human PBMC derived T cell pool. The human T cells were purified from PBMCs using a magnetic bead based separation method, followed by staining with anti-human CD4 (OKT4) mAb conjugated to the PERCP-Cy5.5 fluorophore. The cells were analyzed using flow cytometry. The staining shows two separate populations of CD4+ve and CD4-ve cells, but the separation is not very clear. Also, there is a population with intermediate CD4 expression, which is unusual. The frequencies are lower than physiologically relevant populations and thus, it may not be the best fluorophore to use for surface staining.
Flow Cytometry
Human T cells purified from PBMCs
1:100 dilution for 30 minutes on ice
NA
Flow cytometry on PERCP channel
The staining does separate the positive and negative populations, but not very clearly. There is also some background staining coming up as an intermediate population which makes it difficult to interpret the results. I have used the mAb clone OKT4 several times before for detection of human CD4 and it works pretty well. So, my suggestion is to use a different fluorophore than PERCP for staining.
None
It does the job
The separation is not very clear. Not a reccomended fluorophore.
It works fine for the purpose, but does not produce publication quality data. I would use this only when I have no other choice for a fluorophore.