Hi there - PCR typing will be difficult unless you know the nature of the deletion (for example - if only exon 1 is deleted, then PCRing across eons 2-3 will not be informative..). OF course - you could try Southern Blotting….

Typing by flow cytometry on the other hand will be easy - here is a link to one antibody that can be used, and there may well be others:

https://www.bdbiosciences.com/external_files/pm/doc/tds/brm/live/web_enabled/561183.pdf

Also, testing your knockouts functionally will be really easy - the cells should fail to respond to type-I IFN by STAT1 phosphorylation and will not show up regulation of (for example) MxA or OAS.

Good luck

G

Hi,

Thanks for the suggestions. I looked up the link to find that antibody. However I am a bit puzzled. The antibody is specific to IFNAR1 of course but is also raised in mouse but is unlabelled. So in this case which secondary should I prefer for FACS detection (as using mouse secondary will give background)??

Hi again…

If you can get dissociated cells for flow cytometry (spleen cells, for example) then you don't need to worry about endogenous serum IgG. I'd suggest using any good labelled anti-Mo-IgG1 (or anti-Mo-k if the G1 doesn't work) and go with that - so long as you are sure that your secondary (the antibody we are talking about) does not cross-react with your *blocking* Fc fragments, or Ig mix, it should be fine.

Alternatively, you could biotinylate the antibody yourself (tricky tho with only 0.5 mg)

Good Luck

G

Hi.. Thanks :)

I will try that. I also found the same antibody being sold by BIOLEGEND as PE conjugated.

THANKS again for the help.

PS