Product: Brilliant Violet 421™ anti-mouse CD8a |
Catalog No.: 100737 |
Mikhail Binnewies, Graduate Student UCSF |
Anti-CD8a Brilliant Violet 421™ for Identifying Resident DC Subsets in the LN | |||
Overall: | Product Quality: | Ease of Use: | |
Anti-CD8a Brilliant Violet 421™ staining works well following collagenase I, IV and DNAse digestion in the lymph node. |
Experimental Design |
Application: | Flow cytometry |
Cells used: | Enzymatically digested lymph node cells |
Brief Protocol: | Inguinal lymph nodes were pierced and dissociated in 100 U/ml collagenase I, 400 U/ml collagenase IV and 200 ug/ml DNAse I for 30 minutes at 37C. Digested cells were filtered, washed with DPBS and stained with fixable Zombie NIR™. Cells were washed with PBS/2% FCS/2mM EDTA and stained with a panel to identify T cells, B cells, and DCs, and then anti-CD11b Brilliant Violet 605™ (1:1000) and anti-CD8a Brilliant Violet BV421™ (1:400). Plot shown below is after gating out B and T cells and enriching on resident DCs. |
Results Summary: | Staining of anti-CD8a Brilliant Violet 421™ works very well following enzymatic digestion. |
Additional Notes: | I would not recommend lowering the concentration of the antibody below 1:400. |
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