The polarization state of either pNK cells exposed to PCa cell line (PC-3, DU-145, LNCaP) conditioned media or pNK cells from PCa patients (PCa pTA-NKs), was assessed by flow cytometry for surface antigen expression. Briefly, 2.5 × 105 of total PBMCs per FACS tube were stained for 30 min at 4°C with anti-human monoclonal antibodies (mAbs) as follows: PerCP-conjugated anti-CD3, APC-conjugated anti-CD56, FITC-conjugated anti-CD16, PE-conjugated anti-CD9, PE-conjugated anti-CD49a, PE- conjugated anti-NKG2D, PE-conjugated anti-PD-1, PE- conjugated anti-TIM-3 (all purchased by Miltenyi Biotec). Following Forward/Side Scatter setting, NK cells were identified as CD3- and CD56+ cells (total NK cells). CD16 and NKG2D expression was evaluated on CD3-CD56+ (total NK) gated cells. Finally, CD56 brightness, the expression of the dNK markers CD9, CD49a, expression of CXCR4 and the expression of the exhaustion markers PD-1 and TIM3, were evaluated on total CD3-CD56+NK cells. For details on antibodies used, see Supplementary Table 2
Apc conjugated anti cd56
Manufactured by Miltenyi Biotec
Sourced in United States
The APC-conjugated anti-CD56 is a laboratory reagent used for the detection and analysis of CD56-positive cells in flow cytometry applications. CD56, also known as neural cell adhesion molecule (NCAM), is a cell surface marker expressed on natural killer (NK) cells and a subset of T cells. The APC (Allophycocyanin) fluorescent label allows for the identification and quantification of CD56-positive cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Percp conjugated anti cd3, by Miltenyi Biotec (2 mentions)
Fitc conjugated anti cd16, by Miltenyi Biotec (2 mentions)
Pe conjugated anti cd9, by Miltenyi Biotec (2 mentions)
Pe conjugated anti cd49a, by Miltenyi Biotec (2 mentions)
Pe conjugated anti nkg2d, by Miltenyi Biotec (2 mentions)
Pe conjugated anti pd1, by Miltenyi Biotec (1 mentions)
Fitc conjugated anti cd90, by BD (1 mentions)
Canto 2 instrument, by BD (1 mentions)
Foxp3 staining buffer kit, by Thermo Fisher Scientific (1 mentions)
Pe conjugated anti cd107a, by BD (1 mentions)
Fitc conjugated anti cd44, by BD (1 mentions)
Bv510 conjugated anti cd3, by BD (1 mentions)
Live dead red fluorescent reactive dye, by Thermo Fisher Scientific (1 mentions)
Pe conjugated anti epcam, by BD (1 mentions)
3 protocols using apc conjugated anti cd56
1
Profiling Tumor-Associated NK Cell Phenotypes
2
Multi-Color Flow Cytometry for Cell Characterization
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The following commercially available antibodies were used for multi-color flow cytometry: BV510-conjugated anti-CD3, PE-conjugated anti-CD107a, APC-conjugated anti-ICAM1, PE-conjugated anti-EpCAM, PE-conjugated anti-CD13, FITC-conjugated anti-CD44, FITC-conjugated anti-CD90, PE-conjugated anti-CD133 (BD Biosciences, San Jose, CA, USA), APC-conjugated anti-CD56 (Miltenyi Biotec), APC-conjugated anti-MHC-1, APC-conjugated anti-ULBP-1, APC-conjugated anti-ULBP-2/5/6, APC-conjugated anti-ULBP-3, APC-conjugated anti-TRAIL-R1 (R&D Systems, Minneapolis, MN, USA), APC-conjugated anti-CEACAM1, APC-conjugated anti-HLA-G, and APC-conjugated anti-MICA/B (Biolegend, San Diego, CA, USA). Dead cells were excluded using the LIVE/DEAD red fluorescent reactive dye (Invitrogen). For some experiments, surface marker–stained cells were permeabilized using a Foxp3 Staining Buffer Kit (eBioscience) and further stained for PE-conjugated anti-aldehyde dehydrogenases (ALDH) (Sino Biological, PA, USA). Multi-color flow cytometry was performed using the Canto II instrument (BD Biosciences), and data were analyzed using the FlowJo software (TreeStar, Ashland, OR, USA).
3
NK Cell Phenotyping in Prostate Cancer
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The polarization state of either pNK cells exposed to PCa cell line (PC-3, DU-145, LNCaP) conditioned media or pNK cells from PCa patients, was assessed by flow cytometry for surface antigen expression. Briefly, 2.5x10 5 of total PBMCs per FACS tube were stained for 30 minutes at 4°C with anti-human monoclonal antibodies (mAbs) as follows: PerCP conjugated anti-CD3, APC conjugated anti-CD56, FITC conjugated anti-CD16, PE conjugated anti-CD9, PE conjugated anti-CD49a, PE conjugated anti-NKG2D (all purchased by Miltenyi Biotec). Following Forward/Side Scatter setting, NK cells were identified as CD3 -and CD56 + cells (total NK cells). CD16 and NKG2D expression was evaluated on CD3 -CD56 + (total NK) gated cells. Finally, CD56 brightness and the expression of the dNK markers CD9, CD49a and CXCR4, were evaluated on total CD3 -CD56 + NK cells. For details on antibodies used, see Supplementary table 3.
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