The following mouse monoclonal antibodies were used: CD56-APC (clone N901, Beckman Coulter, Miami, FL, USA), CD56-Brilliant Violet 421 (clone HCD56, Sony Biotechnology, San Jose, CA, USA), CD56-PE (clone N901 (HLDA6), Beckman Coulter, Miami, FL, USA), CD57-PE (clone TB01, eBioscience, San Diego, CA, USA), CD57-FITC (clone TB03, Miltenyi Biotec, Bergisch Gladbach, Germany), CD57-APC (clone TB03, Miltenyi Biotec, Bergisch Gladbach, Germany), CD16-PE (Sorbent, RF), CD2-PE-Cy7 (clone TS1/8, Sony Biotechnology, San Jose, CA, USA), anti-NKG2A-PE (clone 131411, R&D Systems, Minneapolis, MN, USA), anti-KIR2DL2/DL3-PE (clone DX27, Miltenyi Biotec, Bergisch Gladbach, Germany), anti-HLA-DR-FITC (clone B8.12.2, Beckman Coulter, Miami, FL, CA, USA), NKG2A-PE (clone 131411), NKG2C-AlexaFluor488 (clone 108724), NKG2C-PE (clone 134591; R&D Systems, Minneapolis, MN, USA), NKp46-FITC (clone 9E2; Sony Biotechnology, San Jose, CA, USA), anti-NKG2D-PE (clone REA1175, Miltenyi Biotec, Bergisch Gladbach, Germany). Staining was performed according to the manufacturer’s instructions.
Anti nkg2a pe
Manufactured by R&D Systems
Sourced in United States
Anti-NKG2A-PE is a lab equipment product that binds to the NKG2A receptor. It is conjugated with the fluorescent dye phycoerythrin (PE) for detection purposes.
Automatically generated - may contain errors
Lab products found in correlation
Cd56 pe, by Beckman Coulter (2 mentions)
Cd56 apc, by Beckman Coulter (2 mentions)
Anti hla dr fitc, by Beckman Coulter (2 mentions)
Cd57 pe, by Thermo Fisher Scientific (2 mentions)
Cd57 fitc, by Miltenyi Biotec (2 mentions)
Cd57 apc, by Miltenyi Biotec (2 mentions)
Cd2 pe cy7, by Sony (1 mentions)
Nkg2a pe, by R&D Systems (1 mentions)
Nkg2c pe, by R&D Systems (1 mentions)
Anti nkg2d pe, by Miltenyi Biotec (1 mentions)
Anti cd16 pe, by BD (1 mentions)
Anti cxcr3 pe, by BD (1 mentions)
Anti cd19 pe, by BD (1 mentions)
Anti cd14 fitc, by BD (1 mentions)
Anti nkg2c pe, by R&D Systems (1 mentions)
Anti cd3 fluorescein isothiocyanate fitc, by BD (1 mentions)
Anti cd16 phycoerythrin pe, by BD (1 mentions)
Anti dnam 1 pe, by BD (1 mentions)
Anti nkp30 pe, by BD (1 mentions)
Anti nkp44 pe, by BD (1 mentions)
4 protocols using anti nkg2a pe
1
Comprehensive Phenotyping of NK Cells
2
NK Cell Phenotypic Analysis
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The expanded NK cells were stained with the following monoclonal Abs; anti-CD3- fluorescein isothiocyanate (FITC), anti-CD14-FITC, anti-CD16- phycoerythrin (PE), anti-CD19-PE, anti-DNAM-1-PE, anti-CD56-PE-cyanine (Cy)5, anti-CXCR3-PE, anti-NKp30-PE, anti-NKp44-PE, anti-NKp46-PE (all from BD Biosciences, San Jose, CA, USA), anti-NKG2A-PE, anti-NKG2C-PE, and anti-NKG2D-PE (all from R&D systems, Minneapolis, MN, USA). Stained NK cells were acquired on LSR Fortessa and data were analyzed using FlowJo software (TreeStar Inc., Ashland, OR, USA). To confirm phenotypic changes in NK cell receptors in co-culture of tumor cells with NK cells, cryopreserved NK cells were thawed and co-cultured with MIA PaCa-2 at an E:T ratio of 1:1. On 1, 2, or 3 days after co-culture, the harvested NK cells were immunostained with anti-CD16-PE, anti-NKp30-PE, anti-NKp44-PE, anti-NKp46-PE, anti-DNAM-1-PE, anti-CXCR3-PE (all from BD Biosciences, San Jose, CA, USA), anti-NKG2D-PE (R&D systems, Minneapolis, MN, USA), anti-CD96-PE, and anti-CD161-PE (all from eBioscience, San Diego, CA, USA) then analyzed by flow cytometry, as described above.
3
Multiparameter Flow Cytometry of NK Cells
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Flow cytometry of NK cells was performed on a BD LSRII flow cytometer (BD Biosciences, San Jose, CA) and analyzed in FlowJo 9.9 (TreeStar Inc., Ashland, OR). Staining of NK cells was performed with four panels, and the antibodies used were: anti-CD56-BV605, anti-NKG2D-APC, anti-CD107a-PE-Cy7, anti-GranzymeB-FITC, anti-perforin-APC, anti-NKp46-PE-Cy7, anti-NKp30-APC, anti-NKp44-PE, anti-2B4-FITC, anti-4-1BB-PerCP-Cy5.5, anti-CD95-BV421, anti-Tim3-BV421, anti-TRAIL-PE, anti-CD122-PerCP-Cy5.5, anti-CD122-BV510, anti-CD95L-PE, anti-Ki67-BV421, anti-CD40L-BV510, anti-PD-1-PE-Cy7; all were obtained from BioLegend (San Diego, CA). Anti-CD16-APC-H7, PD-L1-PE-Cy7, and CD11a-FITC, were obtained from BD Biosciences (San Jose, CA). Anti-NKG2A-PE was from R&D Systems (Minneapolis, MN), and anti-CD158a-PerCP-Cy5.5 from eBioscience.
4
Phenotyping of Natural Killer Cells
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The following anti-human monoclonal antibodies (mAbs) were used for flow cytometry: CD3- PE-Cy7 (Beckman Coulter, USA, clone UCHT1), CD56-APC (Beckman Coulter, USA, clone N901), CD56-Brilliant Violet 421 (Sony, USA, clone HCD56), CD56-PE (Beckman Coulter, USA, clone N901 (HLDA6)), CD57-PE (eBioscience, USA, clone TB01), CD57-FITC (Miltenyi Biotech, Germany, clone TB03), CD57-APC (Miltenyi Biotech, Germany, TB03), CD16-PE (Sony, USA) anti-NKG2A-PE (R&D Systems, USA, clone 131411), anti-KIR2DL2/DL3-PE (Miltenyi Biotech, Germany, clone DX27), anti-HLA-DR-FITC (Beckman Coulter, USA, clone B8.12.2). Cells were incubated in PBS containing 0.5% BSA (bovine serum albumin) and 0.01% sodium azide (labeling buffer) with mAbs for 30 min on ice, then washed twice with labeling buffer and centrifugation. Samples were subsequently analyzed using FACSCalibur flow cytometer (BD Biosciences, San Jose CA, USA) equipped with 488 and 640 nm lasers. At least 30000 events were recorded in lymphocyte gate for total NK cell population and 5000 events for NK cell clones. Acquired data was analyzed using Flowing Software version 2.5.1 (PerttuTerho, Turku Centre for Biotechnology, Finland) and FlowJo software version 7.6 (FlowJo LLC, Ashland, OR, USA). For fluorescence-activated cell sorting, cells were labeled with mAbs in PBS containing 0.5% BSA and 2 mM EDTA.
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