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2 protocols using chl kir2dl2 3 s2

1

Comprehensive NK Cell Phenotyping

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dNK cells were gated on as live, CD9+CD56+ cells. pbNK cells were gated on as live CD56+CD3 cells. The following Abs were used: Live/Dead discriminator (Life Technologies), CD9 (SN4 or M-L13 from eBioscience or BD Biosciences, respectively), CD56 (HCD56 from BioLegend), and CD3 (SK7) from BD Biosciences. Fibroblasts and macrophages were identified using CD10 (HI10a from BioLegend) and CD14 Abs (MφP9 and HCD14 from BD Pharmingen and BioLegend), respectively. The following Abs were used to stain KIRs: UPR1 (KIR2DL5) from BioLegend and Carlos Vilches (33 (link)); 179315 (KIR2DS4), 143211 (KIR2DL1), and 181703 (KIR2DL4) from R&D Systems; FES172 (KIR2DS4) and EB6 (KIR2DL1/S1) from Beckman Coulter; CHL (KIR2DL2/3/S2) from BD Pharmingen; DX9 (KIR3DL1) from BioLegend; NKVFS1 (KIR2DL1/2/3/S1/2/4) from Abcam; 5.133 (KIR3DL2) from Marco Colonna (34 (link)); and FLAG Abs from Sigma-Aldrich. Intracellular staining was performed according to the manufacturers’ instructions with Abs against Ki647 (BD Pharmingen), CCL3 (R&D Systems), and GM-CSF (BD Biosciences).
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2

Comprehensive NK Cell Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
dNK were gated on as live, CD9+ CD56+ cells. pbNK were gated on as live CD56+ CD3- cells. The following antibodies were used: LIVE/DEAD discriminator (LifeTechnologies), CD9 (SN4 or M-L13 from eBioscience or Becton Dickinson, respectively), CD56 (HCD56 from Biolegend) and CD3 (SK7) from Becton Dickinson. Fibroblasts and macrophages were identified using CD10 (HI10a from Biolegend) and CD14 antibodies (MφP9 and HCD14 from BD Pharmingen and Biolegend), respectively. The following antibodies were used to stain KIR: UPR1 (KIR2DL5) from Biolegend and Carlos Vilches (33 (link)); 179315 (KIR2DS4), 143211 (KIR2DL1) and 181703 (KIR2DL4) from R&D Systems; FES172 (KIR2DS4) and EB6 (KIR2DL1/S1) from Beckman Coulter; CHL (KIR2DL2/3/S2) from BD Pharmingen, DX9 (KIR3DL1) from Biolegend; NKVFS1 (KIR2DL1/2/3/S1/2/4) from Abcam; 5.133 (KIR3DL2) from Marco Colonna (34 ); and FLAG antibodies (Sigma Aldrich). Intracellular staining was performed according to manufacturers’ instructions with antibodies against Ki647, (BD Pharmingen), CCL3 (R&D Systems) and GM-CSF (BD Biosciences).
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