I am doing a multi-color flowcytometry analysis of mouse bone marrow cells, for which I use a kit. Each time, I need only 0.3-0.6 ul of certain antibodies. This has a chance of increasing errors during repetitions. I obtained a huge error bar in my results after N=3. I think it might be due to the pipetting error of the antibodies (probable one of the reasons). Hence, I would like to know if I can prepare a mastermix/bulk mix of the antibodies needed for analysis, stored at 4 degree C and use it at the time of need. Suggestions regarding any other aspect are also welcome !
I try to not use less than 0.5-1.0µl of any one antibody for that reason; I usually prepare a bulk 'cocktail' of antibodies which I then add to each of my samples. I wouldn't store it for longer than a few days at 4 C, since diluting the antibody will also dilute the antimicrobial agent included in the stock. We've had great success in our lab using the protocol for multicolor staining that I've attached to this answer. Good luck!
Hi N. Max Schabla,
Thank you for the suggestions and the protocol.
tandem conjugates e.g. pe-cy5, pe-cy7, apc-cy7 etc, should be added to the mix directly before use as they may split.
Hi Ann,
you are right and I do have a pe-cy7 conjugated antibody included in the staining. Point noted ! Thanks !
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