Hi all,
I'm going to run Flow Cytometery on ferret PBMCs but can't find an Fc block for ferret PBMCs. Does that even exist? Do I need ferret-specific Fc blocking antibodies or can I use: https://www.bdbiosciences.com/us/applications/research/b-cell-research/surface-markers/mouse/purified-rat-anti-mouse-cd16cd32-mouse-bd-fc-block-24g2/p/553142 ? I've seen this used in 2 papers as an Fc block for ferret PBMC flow cytometry and want to make sure it will work fine or if there's a better alternative.
Thanks!
Hi, 2.462 reacts with mouse, human rat, Guinne pig,, and ferret cd16/cd32. I would advise you to also block with ferret serum at the same time. I am assuming you are using direct fluorochrome conjugates in your flow.
Hi, I am using fluorescently-labeled chimeric 44H10 antibody, so yes an direct conjugate. I guess the only way I'll get ferret serum is to use from those same ferrets, haha. But yes, I've had someone suggest to me PBS with inactivated ferret serum as my blocking/staining buffer.
I've been looking for commercial ferret IgG but haven't found any.
Hi my dear, you can purchase normal control ferret serum from genemed synthesis. Its 245$ for 5 mL. Actually, I think this is better than using ferret igg. I'm not sure how many ferrets you have on hand, but if you have to euthanize and your animal protocol allows it, you can collect blood from cardiac puncture to make your own.
I need to do the FACS analysis before we start the experiment actually, so I'd need the ferret serum before we euthanize haha. But thank you for the suggestion from genemed! I'll definitely look into that :)
Hi Lora Benoit so I've decided to take 200uL from one ferret to use the serum as Fc block but I have absolutely no idea of the dilution that I would need to use. Do you know what dilution I should use the serum at in PBS for Fc block?
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