Purified T cells were stimulated as previously described and GM-CSF was measured using the GM-CSF Secretion Assay Enrichment and Detection Kit (PE, Miltenyi, 130-105-760). The manufacturer’s instructions were modified to a 96-well format with final volumes of 200 μl per well. Following the GM-CSF kit protocol, cells were additionally stained with CD4 (clone SK3, BD, BV480, 566104), CD8 (clone SK1, BD, BUV805, 564912), CD3 (clone UCTH1, BD, BB630, 624294), HLA-DR (clone G46-6, BD, BV786, 564041), CD16 (clone 3G8, BD, BV750, 624380), CD45 (clone 2D1, BD, BUV563, 624284), CD69 (clone FN50, BD, APC-H7, 560737), CCR7 (clone G043H7, Biolegend, BV650, 353134), CD45RA (cloneHI100, BD, PE, 561883), CD25 (clone 2A3, BD, PE-Cy7, 335789), CD14 (M5E2, BD, BV570, 624298), CD56 (clone NCAM16.2, BD, PE-Cy5, 624350) and live-dead dye FVS575V (BD, BV570, 565694). Subsequently, cells were fixed and permeabilized in Foxp3/transcription factor staining buffer as previously described, and stained with the above-mentioned metabolic antibodies, before acquiring on the X-30 FACSymphony.
Clone ncam16
Manufactured by BD
Clone NCAM16.2 is a laboratory research tool used to detect and study the expression of the neural cell adhesion molecule (NCAM) in various biological samples. It is a monoclonal antibody that specifically binds to the NCAM16.2 isoform. The core function of this product is to provide a targeted detection method for the NCAM16.2 protein.
Automatically generated - may contain errors
Lab products found in correlation
Clone g043h7, by BioLegend (1 mentions)
Cd3 clone ucth1, by BD (1 mentions)
Clone 2d1, by BD (1 mentions)
Live dead fixable aqua, by Thermo Fisher Scientific (1 mentions)
Anti cd45 apc cy7, by BioLegend (1 mentions)
Clone fn50, by BioLegend (1 mentions)
Novocyte quanteon, by Agilent Technologies (1 mentions)
Anti cd3 pe cy7, by BioLegend (1 mentions)
Clone 2d1, by BioLegend (1 mentions)
2 protocols using clone ncam16
1
Monitoring T cell Activation and Phenotype
2
Assessing NK Cell Activation in Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
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NCI-H1960, A549 and NCI-H1975 cells were seeded in 6-well plates, incubated overnight and treated with docetaxel and cisplatin (concentrations as described above). After 24 hours of treatment, the appropriate conditions were treated with human aCD70 or isotype control and co-cultured with NK cells at a 5:1 ratio. After washing, cells were stained with the following antibodies: anti-CD45 APC-Cy7 (1:50, Clone 2D1, Biolegend), anti-CD3 PE-Cy7 (1:100, Clone SK7, Biolegend), anti-CD56 PE-CF594 (1:25, Clone NCAM16.2, BD Biosciences), anti-CD69 PerCP-Cy5.5 (1:100, Clone FN50, Biolegend) and Live/Dead Fixable Aqua (1:50, Invitrogen) for 30 minutes at 4°C. Acquisition was performed on the Novocyte Quanteon (Agilent technologies).
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