Miltenyi Biotec-tested panel 30 (MBTP 30)
This application protocol describes the analysis of the human KIR repertoire by flow cytometry. An antibody panel is presented that allows the simultaneous assessment of the four major inhibitory KIRs (KIR2DL1, KIR2DL2/DL3/DS2, KIR3DL1) and an additional inhibitory NK cell receptor, namely NKG2A.
Table 1: Antibody panel.
|Specificity||Clone||Purpose||Fluorochrome||Detection filter (nm) (laser)|
|Backbone staining cocktail||7-AAD Staining Solution||-||Viability||-||655-730 (blue)|
|CD3||REA613||T cells||PerCP-Vio® 770||655-730 (blue)|
|CD14||REA599||Monocytes||PerCP-Vio 770||655-730 (blue)|
|CD56||REA196||NK cells||Vio Bright B515||525/50 (blue)|
|Single inhibitory receptor staining cocktail||CD158a (KIR2DL1)||REA284||KIR2DL1+ NK cells||PE/APC*||585/40 (blue)/|
|CD158b (KIR2DL2/DL3)||DX27**||KIR2DL2/DL3+ NK cells||PE/APC*||585/40 (blue)/|
|CD158e (KIR3DL1)||DX9**||KIR2DL1+ NK cells||PE/APC*||585/40 (blue)/|
|CD159a (NKG2A)||REA110||NKG2A+ NK cells||PE/APC*||585/40 (blue)/|
Table 2: Complementary antibody cocktails for single inhibitory receptor stainings.
|Inhibitory receptor of interest||CD158a (KIR2DL1)||CD158b (KIR2DL2/DL3)||CD158e (KIR3DL1)||CD159a (NKG2A)|
|Antibody cocktail for single inhibitory receptor staining||CD158a (KIR2DL1)-PE||CD158b (KIR2DL2/DL3)-PE||CD158e (KIR3DL1)-PE||CD159a (NKG2A)-PE|
|-||CD158a (KIR2DL1)-APC||CD158a (KIR2DL1)-APC||CD158a (KIR2DL1)-APC|
|CD158b (KIR2DL2/DL3)-APC||-||CD158b (KIR2DL2/DL3)-APC||CD158b (KIR2DL2/DL3)-APC|
|CD158e (KIR3DL1)-APC||CD158e (KIR3DL1)-APC||-||CD158e (KIR3DL1)-APC|
|CD159a (NKG2A)-APC||CD159a (NKG2A)-APC||CD159a (NKG2A)-APC||-|
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