We want to separate B cells via FACS, so we need to stain the cells with antibodys, but before we have to incubate the cells with Fc receptor block to avoid false positives.
Our question is how much Fc receptor block we have to use and how many time do we have to incubate the cells with that before adding the antibody cocktail mix.
The Fc receptor block we are going to use is: http://www.bdbiosciences.com/eu/reagents/research/antibodies-buffers/immunology-reagents/anti-mouse-antibodies/cell-surface-antigens/human-bd-fc-block/p/564220
Thanks to everyone!
For murine cells we add ~2 micrograms of anti-FcR to 1x10^6 cells (in 100µl total volume). We incubate for 15 - 30 minutes on ice before adding our staining cocktail. We leave the anti-FcR antibody in the tube while staining (we don't wash it out before adding staining reagents).
We are working with CSF samples from patients, so we don't know how many cells we have. How much anti-FcR should we use then?
Thank you!
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining