iNK cells were incubated for 30 minutes at 37°C with the following monoclonal antibodies: anti-NKG2D (1D11) at 20 μg/ml, anti-NTB-A (NT7) at 5 μg/ml, anti-NKp30 (P30–15) at 10 μg/ml, anti-LFA-1 (HI111) at 10 μg/ml (all from Biolegend), and anti-DNAM-1 at 10 μg/ml (102511) (R&D Systems) alone or in various combinations. iNK cells were then co-cultured with OVCAR8 cells for 4 hours in B0 media and analyzed by flow cytometry for intracellular IFN-γ production using a fluorescently conjugated anti-IFN-γ (B27) (Biolegend) antibody.
Anti nkg2d 1d11
Manufactured by BioLegend
Sourced in United States
Anti-NKG2D (1D11) is a monoclonal antibody that recognizes the NKG2D receptor, a key activating receptor expressed on natural killer (NK) cells, CD8+ T cells, and other immune cell types. This antibody can be used for flow cytometry, immunohistochemistry, and other immunological applications to detect and study NKG2D-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti ifn γ b27, by BioLegend (1 mentions)
Anti ntb a nt7, by BioLegend (1 mentions)
Anti nkp30 p30 15, by BioLegend (1 mentions)
Anti dnam 1, by R&D Systems (1 mentions)
Facs vantage se flow cytometer, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Monensin golgi stop, by BD (1 mentions)
Il 18, by Thermo Fisher Scientific (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Il 15, by Thermo Fisher Scientific (1 mentions)
Anti mr1 clone 26, by BioLegend (1 mentions)
Ghost dye uv450, by Cytek Biosciences (1 mentions)
5 protocols using anti nkg2d 1d11
1
Evaluating iNK Cell Cytotoxicity
2
Flow Cytometric Analysis of iNKT Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The monoclonal antibodies (mAb) used in this study were anti-Vα24Vβ11 (6B11, 10 μL for flow cytometry tests), anti-perforin (dG9), anti-granzyme (CB6), anti-FasL (MFL3), anti-CD107a (1D4B), anti-CD8 (SK1), anti-CD4 (GK1.5), anti-CD3 (17A2), and anti-NKG2d (1D11), which were purchased from Biolegend, Dan Diego, USA (5 μL for flow cytometry tests). For cytofluorimetric analysis, cells were stained with the appropriate mAb, washed, and stained with the appropriate isotype-specific mAb. For intracellular staining, cells were fixed (4% paraformaldehyde), permeabilized (0.1% saponin) (Sigma-Aldrich, Milan, Italy), and stained with the appropriate mAb. All samples were analyzed using a FACSVantage-SE® flow cytometer (BD Biosciences, Milan, Italy). To compare the surface density and the intracellular content of cytotoxic granules among iNKT cells cultured in different conditions, we express them as median fluorescent intensity (MFI). Data analysis was conducted using FACSDiva software (BD Biosciences).
3
Immune Modulation with Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-PVRIG and anti-TIGIT blocking antibodies were provided by Compugen, USA, Inc. Anti-DNAM-1 (11A8), anti-CD16 (3G8), anti-NKp46 (9E2), anti-2B4 (eBioC1.7) and anti-NKG2D (1D11) purified antibodies were purchased from Biolegend. Recombinant human IL-2, IL-12, IL-15 and IL-18 were purchased from Peprotech. Monensin (GolgiStop, BD Biosciences) and brefeldin A (eBioscience) were both used at 1:1,000. Antibodies used for flow cytometry staining are listed in the Online Supplementary Table S1. SKBR3, KG1a, K562, ML-2, THP-1 and Kasumi-1 cell lines were maintained in RPMI 1640 (Gibco) supplemented with Glutamax, penicillin, streptomycin and 10% (or 20% for Kasumi-1) fetal calf serum (FCS). AML-193 cell line was maintained in Iscove's Modified Dulbecco's Media supplemented with 5% FCS, 5 mg/mL transferrin, 5 mg/mL insulin and 2 ng/mL granulocyte-macrophage colony-stimulating factor. All cell lines tested negative for mycoplasma.
4
MAIT Cell-Mediated Breast Cancer Cytotoxicity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell preparations containing primary MAIT cells (CD161+ PBMCs or breast epithelial organoid cells) or in vitro-expanded MAIT cultures were added to wells containing E. coli-exposed or mock-treated MDA-MB-231 breast carcinoma cells. CD161+ PBMCs and in vitro-expanded MAIT cells were added at a 1:1 ratio to breast carcinoma cells, whereas the breast epithelial organoid cells (which are composed of both IELs and epithelial cells) were added at a 3:1 ratio. Where indicated, the following blocking antibodies were added to the cocultures: 20 μg/ml anti-MR1 (clone 26.5; BioLegend) or 5 μg/ml anti-NKG2D (1D11; BioLegend). The carcinoma and effector cells were coincubated at 37 °C for ~ 18 hours, then monensin (GolgiStop) or brefeldin A (GolgiPlug) was added to all cultures, and the cells were coincubated for an additional 6 hours. After ~ 24 hours of coincubation, the effector cells were resuspended using cold EDTA (500 mM) in PBS, washed, and analyzed by flow cytometry.
5
Cytotoxicity Assay of γδ T Cells Against Squamous Cell Carcinoma
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!