I have a primary antibody conjugated with FITC, but the signal is too weak during flow cytometry. Would it be possible to use an FITC labeling kit to relabel the antibody and enhance the signal? Or is there another way to improve the signal? Even after titrating the antibody to a concentration of 1:20, the signal is still very low—lower than the IgG control.
I also have another unconjugated antibody, but its concentration is 0.2 mg/mL, which is lower than the recommended concentration of 0.5 mg/mL for the labeling kit. Has anyone tried something like this before?
Dear Jiyoon Kim
if the buffer of your labelled antibody is compatible with the kit, you can certanilly try to re-label it.
If you are using a randomic labelling strategu (eg labelling on the lysines) just check that the bbinding of the antibody to the antigen is not affected by labelling, since there is ever a small risk that the FITC label lysisne in the CDR and reduce the antibody binding capability.
best
Manuele
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