In this test, human in vitro-expanded CD8 cells or total murine-PBMCs were incubated with target human or mouse tumor cells, respectively. Anti-CD107a monoclonal antibody conjugated with APC (BD), brefeldin A 10 μg/ml (BD) and GolgiStop containing monensin (BD), the HLA-I pan blocker as control (10 ug/ml, Clone W6/32, BioXcell) were added to the cell culture for a final volume of 200 μl and incubated for 5 h at 37°C. Cells were then stained with anti-CD3a, anti-CD8, and anti-CD4 (BD) antibodies. Intracellular staining for IFN-γ (BD) production was also performed in order to further assess T cell activation.
Clone w6 32
Manufactured by BioXCell
The Clone W6/32 is a monoclonal antibody that recognizes the human major histocompatibility complex (MHC) class I molecule. It is commonly used in various laboratory applications, such as flow cytometry and immunohistochemistry, to detect and study MHC class I expression on cells.
Automatically generated - may contain errors
2 protocols using clone w6 32
1
CD8 T Cell Cytotoxicity Assay
2
Antigen-Specific CTL Activation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human acute monocytic leukemia Thp1 cell line was loaded either with antigens at different concentrations (20 uM-1.25 uM) or with conditioned medium and used to stimulate CTL-Vax (ratio 1:1). Cells’ supernatant was collected after 72 hours and IFN-γ production was measured by ELISA (R&D). IFN-γ released by CTLs cocultured with unloaded Thp1 (spontaneous IFN-γ release) was subtracted from IFNγ measured upon antigen stimulation. Activation specificity was further assessed using an HLA-I pan blocker as control (10 μg/ml, Clone W6/32, BioXcell).
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