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Category 'Antibodies'
Status: Unanswered
Views: 172

IL22 Clone for staining human CD3+CD4+ cells- difficulty staining

fiona cunningham (Author)

Friday, April 12, 2019 4:18 PM

Dear all,

I am most grateful for advice on the best IL22 clone to use for staining CD3+CD4+ lymphocytes using flow cytometry. 

I am having some difficulty with clone 142928 which I happen to have used as conjugated with PE for the panel.

I am not sure if it is a clone issue with IL22 or the problem is with the spectral overlap with BV605-IL17A antibody. 


thanks in advance,

Fiona.

Lynda Guzik

Friday, April 12, 2019 7:30 PM

Hi Fiona,

You don't say which instrument you are using or what the rest of your panel looks like.  However, BV605 and PE, even though they are on different laser lines, contribute a lot of overlap and spreading error to each other.  In addition, IL22 and IL17 are coexpressed by the same cells.  So, my guess is a panel design problem.  One way to prove that would be with 2 FMO controls - one without the IL22 and the other without the IL17.  If you get decent results in those tubes, it's time to consider moving one of those two ab's.  Good luck!

Lynda