We are currently trying to set up a protocol to isolate immune cells from mouse epidermis and dermis for analysis of immune cells by FACS. Dispase works very well to separate the 2 layers. However most of the usual T and B cell markers are destroyed and not recognized anymore by the FACS antibodies. Any suggestion about the type of dispase to use or an alternative to dispase?
Thank you very much for your help.
Best regards
We use Dispase II from Sigma and it works very well at 2 mg/mL.
Dispase would not disturb the expression of lymphocyte surface markers as far as we have tested.
Thank you very much Fangzhou for your very rapid answer.
How long do you expose the mouse skin to dispase?
I want to ask a question. Recently, I also started my work on analysis of immune cells in the skin by FACS. But what i'm confused is that when I cut off the skin from the back of mouse, the skin can apparently be distinguished into two layers, the outer one is tight and white while the inner one is floppy. SO, is the dermis just the floppy one or is the dermis a part of the tight white layer?
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