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Anti nkg2d or rat igg1 isotype antibody

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Anti-NKG2D or rat IgG1 isotype antibody is a laboratory reagent used in immunological research. It functions as a tool to detect or study the NKG2D receptor, which is expressed on the surface of natural killer cells and some T cells. The antibody can be used in various immunoassays and flow cytometry applications.

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Lab products found in correlation

Golgiplug, by BD (2 mentions) Golgistop, by BD (2 mentions) Anti ifnγr1 or rat igg2a isotype antibody, by Thermo Fisher Scientific (2 mentions) Anti mhcii or rat igg2b isotype antibody, by BioXCell (2 mentions) Celltrace violet ctv dye, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

Rat IgG1 (14 citations) Anti-NKG2D (14 citations)

4 protocols using anti nkg2d or rat igg1 isotype antibody

1

Tumor-Immune Cell Co-culture Assay

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NK and CD4+ T cells were isolated from spleens of mice harboring control tumors and enriched as described above. Tumor cells (Control or Rnf2 KO) were enriched as CD45− cells as described above, followed by co-culture in triplicates with CD4+ T cells, NK cells or CD4+ T cells plus NK cells (tumor: immune = 1:5) for 16 h in the presence of hIL-2 (50 U/ml) and soluble anti-CD28 (2 μg/ml, for groups with CD4+ T cells). Cells were then incubated with the Golgi Plug (BD Biosciences) plus Golgi Stop (BD Biosciences) for 5 h prior to flow cytometry analysis of IFNγ production by CD4+ T cells or NK cells. In some groups, 30 μg/ml anti-NKG2D or rat IgG1 isotype antibody (Biolegend) was pre-incubated with NK cells or CD4+ T cells for 15 min before addition to the co-culture, and 10 μg/ml anti-IFNγR1 or rat IgG2a isotype antibody (Fisher) was pre-incubated with NK cells or CD4+ T cells for 30 min before addition to the co-culture. 10 μg/mL anti-MHCII or rat IgG2b isotype antibody (BioXcell), or 10 μg/mL antibodies to IFNγ, IL-2 or recombinant mouse NKG2D Fc (blocking NKG2DL) were added into some groups. Groups with CD4+ T cells or NK cell alone were also included for analysis.
Zhang Z., Luo L., Xing C., Chen Y., Xu P., Li M., Zeng L., Li C., Ghosh S., Manna D.D., Townes T., Britt W.J., Wajapeyee N., Sleckman B.P., Chong Z., Wu Leavenworth J, & Yang E.S. (2021). RNF2 ablation reprograms the tumor-immune microenvironment and stimulates durable NK and CD4+ T-cell-dependent antitumor immunity. Nature cancer, 2(10), 1018-1038.
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2

Cytotoxicity Assay of Engineered Tumor Cells

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Enriched CD4+ T cells were stimulated with plate-coated anti-CD3 antibody (5 μg/ml) and soluble anti-CD28 antibody (2 μg/ml) in the presence of recombinant human IL-2 (hIL-2) (50 U/ml) for 72 h. Enriched NK cells were incubated with hIL-2 (500 U/ml) for 72 h. Enriched tumor cells (Control or Rnf2 KO) were labeled with CellTrace Violet (CTV) dye (5 μM) according to the manufacturer’s protocol (ThermoFisher Scientific) followed by co-culture in triplicates without or with activated CD4+ T cells, NK cells or CD4+ T cells plus NK cells (the ratio of tumor: immune effector cells = 1:5) in the presence of hIL-2 (50 U/ml) for 16 h prior to staining with the fixable viability dye and flow cytometry analysis. In some cases, 30 μg/ml anti-NKG2D or rat IgG1 isotype antibody (Biolegend) was pre-incubated with NK cells or CD4+ T cells for 15 min before addition to the co-culture. Cells positive for both CTV and fixable viability dye are defined as dead cells. The percent tumor killing is calculated using the following equation: [%dead cells (experimental group)-% dead cells (tumor alone)]/% dead cells (tumor alone).
Zhang Z., Luo L., Xing C., Chen Y., Xu P., Li M., Zeng L., Li C., Ghosh S., Manna D.D., Townes T., Britt W.J., Wajapeyee N., Sleckman B.P., Chong Z., Wu Leavenworth J, & Yang E.S. (2021). RNF2 ablation reprograms the tumor-immune microenvironment and stimulates durable NK and CD4+ T-cell-dependent antitumor immunity. Nature cancer, 2(10), 1018-1038.
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3

Cytotoxicity Assay of Engineered Tumor Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Enriched CD4+ T cells were stimulated with plate-coated anti-CD3 antibody (5 μg/ml) and soluble anti-CD28 antibody (2 μg/ml) in the presence of recombinant human IL-2 (hIL-2) (50 U/ml) for 72 h. Enriched NK cells were incubated with hIL-2 (500 U/ml) for 72 h. Enriched tumor cells (Control or Rnf2 KO) were labeled with CellTrace Violet (CTV) dye (5 μM) according to the manufacturer’s protocol (ThermoFisher Scientific) followed by co-culture in triplicates without or with activated CD4+ T cells, NK cells or CD4+ T cells plus NK cells (the ratio of tumor: immune effector cells = 1:5) in the presence of hIL-2 (50 U/ml) for 16 h prior to staining with the fixable viability dye and flow cytometry analysis. In some cases, 30 μg/ml anti-NKG2D or rat IgG1 isotype antibody (Biolegend) was pre-incubated with NK cells or CD4+ T cells for 15 min before addition to the co-culture. Cells positive for both CTV and fixable viability dye are defined as dead cells. The percent tumor killing is calculated using the following equation: [%dead cells (experimental group)-% dead cells (tumor alone)]/% dead cells (tumor alone).
Zhang Z., Luo L., Xing C., Chen Y., Xu P., Li M., Zeng L., Li C., Ghosh S., Manna D.D., Townes T., Britt W.J., Wajapeyee N., Sleckman B.P., Chong Z., Wu Leavenworth J, & Yang E.S. (2021). RNF2 ablation reprograms the tumor-immune microenvironment and stimulates durable NK and CD4+ T-cell-dependent antitumor immunity. Nature cancer, 2(10), 1018-1038.
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4

Tumor-Immune Cell Co-culture Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
NK and CD4+ T cells were isolated from spleens of mice harboring control tumors and enriched as described above. Tumor cells (Control or Rnf2 KO) were enriched as CD45− cells as described above, followed by co-culture in triplicates with CD4+ T cells, NK cells or CD4+ T cells plus NK cells (tumor: immune = 1:5) for 16 h in the presence of hIL-2 (50 U/ml) and soluble anti-CD28 (2 μg/ml, for groups with CD4+ T cells). Cells were then incubated with the Golgi Plug (BD Biosciences) plus Golgi Stop (BD Biosciences) for 5 h prior to flow cytometry analysis of IFNγ production by CD4+ T cells or NK cells. In some groups, 30 μg/ml anti-NKG2D or rat IgG1 isotype antibody (Biolegend) was pre-incubated with NK cells or CD4+ T cells for 15 min before addition to the co-culture, and 10 μg/ml anti-IFNγR1 or rat IgG2a isotype antibody (Fisher) was pre-incubated with NK cells or CD4+ T cells for 30 min before addition to the co-culture. 10 μg/mL anti-MHCII or rat IgG2b isotype antibody (BioXcell), or 10 μg/mL antibodies to IFNγ, IL-2 or recombinant mouse NKG2D Fc (blocking NKG2DL) were added into some groups. Groups with CD4+ T cells or NK cell alone were also included for analysis.
Zhang Z., Luo L., Xing C., Chen Y., Xu P., Li M., Zeng L., Li C., Ghosh S., Manna D.D., Townes T., Britt W.J., Wajapeyee N., Sleckman B.P., Chong Z., Wu Leavenworth J, & Yang E.S. (2021). RNF2 ablation reprograms the tumor-immune microenvironment and stimulates durable NK and CD4+ T-cell-dependent antitumor immunity. Nature cancer, 2(10), 1018-1038.
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