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Cd1d tetramer

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The CD1d tetramer is a laboratory reagent used for the identification and characterization of CD1d-restricted T cells. It consists of recombinant CD1d molecules complexed with a specific lipid antigen, allowing for the detection and enumeration of CD1d-reactive T cells in biological samples.

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Lab products found in correlation

Cytofix cytoperm, by BD (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions) Perm wash buffer, by BD (2 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Anti cd16 32, by BioLegend (1 mentions) Cytofix cytoperm solution, by Thermo Fisher Scientific (1 mentions) Mp4 25d2, by BioLegend (1 mentions) Tcrβ h57 597, by BioLegend (1 mentions) Ki67 sola15, by Thermo Fisher Scientific (1 mentions) Cd45 hi30, by Thermo Fisher Scientific (1 mentions) Cd69 h1.2f3, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm solution, by BD (1 mentions) Cd69 fn50, by BioLegend (1 mentions) Cd45 30 f11, by BioLegend (1 mentions)

Close spelling variants of this product

CD1d-PBS57 tetramers (1 citations)

3 protocols using cd1d tetramer

1

Comprehensive Immune Cell Profiling

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Single cell suspensions were stained with fixable viability dye eFluor® 780 (eBioscience) for 30 min at 4°C, and Fc receptors were blocked with anti-CD16/32 (BioLegend) blocking antibody prior to surface staining with antibodies. Antibodies for surface staining used are listed, mouse: CD45 (30-F11, BioLegend), TCR β (H57-597, BioLegend), CD69 (H1.2F3, eBioscience), CD1d tetramer (NIH); human: CD45 (HI30, eBioscience), TCRα/β (IP26, BioLegend), CD69 (FN50, BioLegend), Vα24 (6B11, BD Bioscience). For intracellular staining, single cell suspensions were stimulated with 50 ng/mL phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich), 1 μg/mL ionomycin (Sigma-Aldrich) and 1 μg/mL Golgi stop A (BD Biosciences) for 4 h. After surface staining, cells were fixed and permeabilized with Cytofix/Cytoperm solution (BD biosciences) and further stained intracellularly with human IFNγ (B27, BioLegend) and human IL-4 (MP4-25D2, BioLegend). For Ki67 staining, cells were fixed and permeabilized with Cytofix/Cytoperm solution (eBioscience), and further stained with Ki67 (SolA15, eBioscience). Data were acquired using LSR II (BD Biosciences) and analyzed using the FlowJo software (BD Biosciences).
Lai A.C., Chi P.Y., Thio C.L., Han Y.C., Kao H.N., Hsieh H.W., Gervay-Hague J, & Chang Y.J. (2019). α-Lactosylceramide Protects Against iNKT-Mediated Murine Airway Hyperreactivity and Liver Injury Through Competitive Inhibition of Cd1d Binding. Frontiers in Chemistry, 7, 811.
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2

Intracellular Cytokine and Transcription Factor Staining

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To stain cytokines intracellularly, cells were fixed and permeabilized with BD Cytofix/Cytoperm followed by staining with fluorescently labeled antibodies against IFN-γ, IL-4, or IL-17 in Perm wash buffer (BD Biosciences). To analyze expression of transcription factors in iNKT cells, thymocytes were stained with fluorescently conjugated CD1d tetramer, followed by fixation/permeabilization using the Foxp3/transcription factor staining buffer set (eBioscience). Subsequently, cells were incubated with purified anti-PLZF, followed by staining with fluorescently labeled anti-mouse IgG1. Thereafter, cells were stained with antibodies against T-bet and Rorγt.
Beyaz S., Kim J.H., Pinello L., Xifaras M.E., Hu Y., Huang J., Kerenyi M.A., Das P.P., Barnitz R.A., Herault A., Dogum R., Haining W.N., Yilmaz Ö.H., Passegue E., Yuan G.C., Orkin S.H, & Winau F. (2016). The histone demethylase UTX regulates the lineage-specific epigenetic program of invariant natural killer T cells. Nature immunology, 18(2), 184-195.
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3

Intracellular Cytokine and Transcription Factor Staining

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To stain cytokines intracellularly, cells were fixed and permeabilized with BD Cytofix/Cytoperm followed by staining with fluorescently labeled antibodies against IFN-γ, IL-4, or IL-17 in Perm wash buffer (BD Biosciences). To analyze expression of transcription factors in iNKT cells, thymocytes were stained with fluorescently conjugated CD1d tetramer, followed by fixation/permeabilization using the Foxp3/transcription factor staining buffer set (eBioscience). Subsequently, cells were incubated with purified anti-PLZF, followed by staining with fluorescently labeled anti-mouse IgG1. Thereafter, cells were stained with antibodies against T-bet and Rorγt.
Beyaz S., Kim J.H., Pinello L., Xifaras M.E., Hu Y., Huang J., Kerenyi M.A., Das P.P., Barnitz R.A., Herault A., Dogum R., Haining W.N., Yilmaz Ö.H., Passegue E., Yuan G.C., Orkin S.H, & Winau F. (2016). The histone demethylase UTX regulates the lineage-specific epigenetic program of invariant natural killer T cells. Nature immunology, 18(2), 184-195.
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