Peripheral blood was collected from healthy donors using protocols approved by the Memorial Sloan Kettering Cancer Center Institutional Review Board (nos. 06-107 and 95-054). Specific processing on these samples was performed under Biospecimen Research Protocol Institutional Review Board no. 16-1564. Donors provided informed written consent. PBMCs were isolated by Ficoll gradient purification. Flow cytometry and cell sorting were performed on the LSR II and Aria II cytometers (BD Biosciences), respectively. NK cells were sorted and/or phenotyped from PBMCs by using the following fluorophore-conjugated antibodies at a 1:50 dilution, unless otherwise indicated: DAPI, CD3ε (1:200 dilution, UCHT1, BD Biosciences), CD56 (1:100 dilution, N901, Beckman Coulter) and CD14 (M5E2, BD Biosciences), CD57 (HCD57, BioLegend) and NKG2C (134591, R&D). Cells were sorted to >95% purity.
Cd57 hcd57
Manufactured by BioLegend
CD57 (HCD57) is a cell surface glycoprotein expressed on natural killer (NK) cells, a subset of T cells, and some monocytes. It is a marker for late-stage or terminally differentiated NK cells and T cells.
Automatically generated - may contain errors
Lab products found in correlation
Cd14 m5e2, by BD (1 mentions)
Nkg2c, by R&D Systems (1 mentions)
Aria 2, by BD (1 mentions)
Cd16 3g8, by BD (1 mentions)
Perforin δg9, by BD (1 mentions)
Granzyme b gb11, by BD (1 mentions)
Granzyme a cb9, by BD (1 mentions)
Cd56 ncam16.2, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Cd45 30 f11, by Thermo Fisher Scientific (1 mentions)
2 protocols using cd57 hcd57
1
Isolation and Phenotyping of Human NK Cells
2
Comprehensive Immune Cell Profiling
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For cell staining and flow cytometry, fluorochrome-conjugated antibodies to CD3 (clone S4.1; Invitrogen), CD4 (S3.5; Invitrogen), CD8 (RPA-T8; BioLegend), CD16 (3G8; BD Biosciences), CD45 (30-F11; Invitrogen), CD56 (NCAM16.2; BD Biosciences), CD57 (HCD57; BioLegend), CD107a (H4A3; BD Biosciences), perforin (δG9; BD Biosciences), granzyme A (CB9; BD Biosciences), and granzyme B (GB11; BD Biosciences), were used. Fluorochrome-conjugated IgG1 and IgG2b (MOPC-21 and 27-35; BD Biosciences) isotype control antibodies were also used in addition to a fixable live/dead cell marker (Invitrogen). Mouse anti-CD3 (S4.1) and purified anti-CD16 (3G8) mAbs were used for redirected ADCC. For confocal microscopy, mouse monoclonal antibodies to α-tubulin (236-10501; Invitrogen), mannose-6-phosphate receptor (MEM-238; Invitrogen), and perforin (δG9; BioLegend) mAbs were used. Rabbit polyclonal antibodies used were Rab27a, Stx11, and Munc13-4 (all Protein Technologies Group), EEA1 (Cell Signaling Technologies), WASP-interacting protein (WIP, Santa-Cruz), and Cathepsin D (Upstate). Secondary antibodies were donkey anti-mouse and donkey anti-rabbit (both Invitrogen). DNA was labeled with DAPI and actin with phalloidin (both Invitrogen).
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