To analyze the expression of NK cell receptors on NK-92 cells, the NK-92 cells were stained with the following antibodies: PE-conjugated anti-NKP30 (Catalog # 558407, BD Biosciences), PE-conjugated anti-NKP46 (Catalog # 557991, BD Biosciences), PE-conjugated anti-CD158a (Catalog # 556063, BD Biosciences), PE-conjugated anti-NKG2A (Catalog # FAB1059P, R&D Systems), PE-conjugated anti-NKG2D (Catalog # FAB139P, R&D Systems), PE-conjugated lgG1 isotype control (Catalog # 556063, BD Biosciences), and PE-conjugated lgG2A isotype control (Catalog # IC003P, R&D Systems). Intracellular cytokine staining analysis was used to detect the expression of IL-32 in NK-92 cells, liver NK cells and T cells. In brief, NK-92 cells were stimulated by directly adding an anti-NKP30 monoclonal antibody (Catalog # MAB18491, Clone 210847, R&D Systems) and IgG2A Isotype Control (Catalog # 20102, R&D Systems) with final concentration of 1ug/ml or co-culturing with Huh7 cells in the presence of BD GolgiStop Protein Transport inhibitor. Liver NK cells and T cells were stimulated by phorbol myristate acetate (PMA) and inomycin in the presence of BD GolgiStop Protein Transport inhibitor. After stimulation for 4h, cells were washed, fixed, permeabilized and stained with APC-conjugated anti-IL-32 (Catalog # IC30402A, R&D Systems) and APC-conjugated lgG2A isotype control (Catalog # IC006A, R&D Systems).
Pe conjugated anti nkp30
Manufactured by BD
PE-conjugated anti-NKP30 is a laboratory reagent for the detection and analysis of NKP30 expression on cell surfaces. It is a fluorochrome-labeled antibody that binds specifically to the NKP30 antigen, allowing for its identification and quantification using flow cytometry or other compatible techniques.
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2 protocols using pe conjugated anti nkp30
1
NK Cell Receptor Expression and Cytokine Analysis
2
NK-cell Phenotyping and Ligand Analysis
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For NK-cell phenotyping, 2.0 X10 5 PBMC were incubated with the appropriated Abs: FITC-conjugated anti-CD3, allophycocyanin-conjugated anti-CD56 and anti-CD69, PE-conjugated anti-NKp30, anti-NKp44, anti-NKp46, anti-CD16, anti DNAM-1, anti-CD94, anti-CD161, anti-CD56, anti-CD158a/h, anti-CD158b, anti-CD107a and anti-CD85j (BD PharMingen) or PE-conjugated anti-NKG2A (R&D Systems), for 30 min at 4°C. The NK-cell population was selected based on CD3 -CD56 + phenotype of gated lymphocytes according to forward-side scatter. For NK-cell ligand analysis, tumor cell lines were incubated with FITC-conjugated anti-HLA-ABC (BD PharMingen), or PE-conjugated anti-HLA-G, anti-HLA-E (Abcam), anti-CD112 and ant-MICA/B (BD PharMingen), anti-CD155 (BioLegend) or respective isotype controls. All samples were acquired on a BD FACS Calibur using Cellquest Pro software (BD Biosciences) and analyzed with FlowJo 7.6.2 software (Tree Star, Inc.).
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