Titrating the antibodies

Dear All,

Is it necessary to titrate all the antibodies before doing the FACS
analysis, eventhough most of my antibodies are pre-titrated/pre-diluted (as
per datasheet) but some of them say only concentration like 0.2 mg. I have
got 15 antibodies and would appreciate any help/advice in this regard. Can
anyone please suggest the best way to titer the antibodies, I will be doing
3 color flow cytometry.

I would also like to know if it is best to buy the antibody and isotype
control from the same company ? I bought one antibody from a different
company but I already have the isotype control from another company, now is
it recommended and worth buying the isotype control from the same company
from where I bought the antibody or shall I buy one ?

I shall look forward to your responses. Thanks a lot.


Best wishes,

Abdul