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Category 'Antibodies'
Status: Unanswered
Views: 246

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,


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!