The following anti-human antibodies were used in this study: anti-CD3-BV605 (clone OKT3, Biolegend), anti-CD3-BUV395 (clone UCHT1, BD Bioscience), anti-CD14-V500 (clone M5E2, BD Bioscience), anti-CD19-BV510 (clone SJ25C1, BD Bioscience), anti-CD56-PE-Cy7 (clone NCAM16.2, BD Bioscience), anti-CXCR6-PE or anti-CXCR6-APC (clone K041E5, Biolegend), anti-CD57-AF488 (clone TB01, eBioscience), anti-TRAIL-BV421 (clone RIK-2, BD Bioscience), anti-CD49a-FITC (clone TS2/7, Biolegend), anti-HLA-DR V500 (clone G46-6, BD Bioscience) and anti-CD69-PE-Dazzle (clone FN50, Biolegend) for surface antigens; and anti-T-bet-eFlour610 (clone 4B10, eBioscience) and anti-Eomes-PE-eFlour610 (clone Dan11mag, eBioscience) for intranuclear antigens; anti-IFNγ-V450 (BD Bioscience), anti-MIP-1β-PerCP-Cy5.5 (cloneD21–1351, BD Bioscience), anti-GM-CSF-PE-CF594 (clone BVD2–21C11, BD Bioscience), anti-TNF-FITC (clone MAb11, BD Bioscience), anti-Granzyme B-AlexaFlour700 (clone GB11, BD Bioscience) and anti-Perforin-PerCP-Cy5.5 (clone dG9, Biolegend) for intracellular antigens.
Anti cd56 pe cy7 clone ncam16
Manufactured by BD
Anti-CD56-PE-Cy7 (clone NCAM16.2) is a fluorochrome-conjugated monoclonal antibody that recognizes the CD56 antigen, also known as the neural cell adhesion molecule (NCAM). This antibody is designed for flow cytometric analysis and can be used to identify and enumerate CD56-positive cells within a sample.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd3 bv605 clone okt3, by BioLegend (1 mentions)
Anti mip 1β percp cy5, by BD (1 mentions)
Anti cd3 buv395 clone ucht1, by BD (1 mentions)
Anti ifn γ v450, by BD (1 mentions)
Anti cd4, by Miltenyi Biotec (1 mentions)
Anti cd8 fitc clone rpat8, by BD (1 mentions)
Anti cd11c apc clone b ly6, by BD (1 mentions)
Anti cd14 pb clone m5e2, by BD (1 mentions)
Ficoll, by GE Healthcare (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
Anti cd56 microbeads, by Miltenyi Biotec (1 mentions)
Anti cd3 percp clone sk7, by BD (1 mentions)
Facs lysis solution, by BD (1 mentions)
Anti nkg2c pe clone 134591, by R&D Systems (1 mentions)
Live dead blue viability dye, by Thermo Fisher Scientific (1 mentions)
Anti nkg2a apc clone z199, by Beckman Coulter (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
3 protocols using anti cd56 pe cy7 clone ncam16
1
Comprehensive Immune Profiling using Multicolor Flow Cytometry
2
PBMC Isolation and Cell Sorting
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Peripheral blood mononuclear cells (PBMC) were freshly isolated by Ficoll (GE Healthcare Biosciences) as described previously [37 (link)]. PBMC subpopulations were sorted with anti-CD4, anti-CD8, anti-CD14, anti-CD19, and anti-CD56 MicroBeads (Miltenyi Biotec) with an autoMACS Pro Separator (Miltenyi Biotec) according to manufacturer’s instructions. The purity of sorted cells was checked by flow cytometry with the following antibodies: anti-CD4 APC-H7 (clone SK3, BD Biosciences), anti-CD4 ECD (clone SFCI12T4D11, Beckman Coulter), anti-CD8 FITC (clone RPA-T8, BD Biosciences), anti-CD11c APC (clone B-ly6, BD Biosciences), anti-CD14 PB (clone M5E2, BD Biosciences), anti-CD19 PE (clone HIB19, eBioscience), and anti-CD56 PE-Cy7 (clone NCAM16.2, BD Biosciences) on a LSRII flow cytometer (BD Biosciences). We did not pursue the proposed experiments if the purity of sorted cells was less than 90 %. Analyses were performed using FlowJo software (Treestar).
3
NK Cell Phenotyping in HIV Infection
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To assess the expression of CD57, NKG2A and NKG2C on NK cells from HIV-1-infected and uninfected donors, 200 μl whole blood or 10 6 PBMC were stained with the following antibodies for 30 minutes at room temperature: anti-CD3 PerCP (clone SK7; BD Biosciences) or anti-CD3 BV785 (clone SK7; Biolegend), anti-CD56 PE-Cy7 (clone NCAM16.2; BD Biosciences), anti-CD57 Pacific Blue (clone HCD57; Biolegend), anti-NKG2C PE (clone 134591; R&D Systems) and anti-NKG2A APC (clone Z199; Beckman Coulter). After whole blood staining, red blood cells were lysed in BD FACS lysis solution (BD Biosciences) and cells were washed and fixed in 1% formaldehyde. After PBMC staining, cells were washed and fixed. Samples were acquired on an LSR Fortessa flow cytometer (BD Biosciences). Flow cytometry data were analyzed using FlowJo software (FlowJo LLC). For analysis of the expression of NKG2A on CD57 -, CD57 + NKG2C -and CD57 + NKG2C + NK cell subsets, donors with less than 100 total events in any of the three NK cell subsets were excluded from analysis.
Staining of pre-and post-ART cryopreserved samples from the IVRN was performed as described above for fresh samples, with the additional inclusion of a Live/Dead blue viability dye (Thermo Fisher).
Staining of pre-and post-ART cryopreserved samples from the IVRN was performed as described above for fresh samples, with the additional inclusion of a Live/Dead blue viability dye (Thermo Fisher).
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