CD8+ T-cells were isolated using the MagCellect Mouse CD8+ T Cell Isolation Kit (R&D Systems) from the spleens and lymph nodes of naive Balb/c mice. Purified CD8+ T-cells were placed into culture and stimulated with anti-CD3 (2μg/mL) antibody for 1 h prior to transfer onto purified primary microglial or astrocyte cell cultures with or without IFN-γ pretreatment (8 h; 200U/mL; eBioscience). CD8+ T-cells were added at a 10:1 CD8: glial cell ratio. Neutralization of PD-1 and its ligand was performed by treating glial cells with anti-PD-1 (J43 clone; eBiosciences), anti-PD-L1 (M1H5 clone; eBiosciences), anti-PD-L2 (TY25 clone; eBiosciences), or IgG2a for 2 h prior to the addition of anti-CD3-activated CD8 T-cells. Supernatants were collected 48 h after the addition of T-cells and frozen at −20°C until analysis. IFN-γ and IL-2 concentration were measured using a mouse IFN-γ or IL-2 ELISA kit (eBioscience) according to manufacturers instructions.
Mouse ifn γ or il 2 elisa kit
Manufactured by Thermo Fisher Scientific
The Mouse IFN-γ or IL-2 ELISA kit is a laboratory tool used to detect and quantify the presence of interferon-gamma (IFN-γ) or interleukin-2 (IL-2) in mouse samples. This kit employs the enzyme-linked immunosorbent assay (ELISA) technique to measure the analyte of interest.
Automatically generated - may contain errors
Lab products found in correlation
Anti pd 1 clone j43, by Thermo Fisher Scientific (2 mentions)
Ifn γ, by Thermo Fisher Scientific (2 mentions)
Anti pd l1 m1h5 clone, by Thermo Fisher Scientific (2 mentions)
Anti pd l2 ty25 clone, by Thermo Fisher Scientific (2 mentions)
Magcellect mouse cd8 t cell isolation kit, by R&D Systems (2 mentions)
2 protocols using mouse ifn γ or il 2 elisa kit
1
Activated CD8+ T-Cell Interaction with Glia
2
CD8+ T-Cell Activation and Cytokine Release
Check if the same lab product or an alternative is used in the 5 most similar protocols
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CD8+ T‐cells were isolated using the MagCellect Mouse CD8+ T Cell Isolation Kit (R&D Systems) from the spleens and lymph nodes of naive Balb/c mice. Purified CD8+ T‐cells were placed into culture and stimulated with anti‐CD3 (2 µg/mL) antibody for 1 h prior to transfer onto purified primary microglial or astrocyte cell cultures with or without IFN‐γ pretreatment (8 h; 200U/mL; eBioscience). CD8+ T‐cells were added at a 10:1 CD8: glial cell ratio. Neutralization of PD‐1 and its ligand was performed by treating glial cells with anti‐PD‐1 (J43 clone; eBiosciences), anti‐PD‐L1 (M1H5 clone; eBiosciences), anti‐PD‐L2 (TY25 clone; eBiosciences), or IgG2a for 2 h prior to the addition of anti‐CD3‐activated CD8 T‐cells. Supernatants were collected 48 h after the addition of T‐cells and frozen at −20°C until analysis. IFN‐γ and IL‐2 concentrations were measured using a mouse IFN‐γ or IL‐2 ELISA kit (eBioscience) according to manufacturer's instructions.
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