The organoids were labeled with Green Dye (Thermo Fisher, C7025), and organoid-primed T (opT) cells were added at a ratio of 1:10 (Tumor cell: T cell). Images were taken every 10 min starting at 2.0-hour co-culture for a total of 20 hours. For killing assay, opT cells were added to three-dimensional (3D) organoids at a ratio of 1:1 and incubated at 37°C for 24 hours, 48 hours, and 72 hours, respectively. Supernatants were collected and used for ELISA: M30 Apoptosense CK18 Kit (DiaPharma, P10011), interferon (IFN)-γ ELISA kit (Mabtech, 3420–1 H-20), and granzyme B ELISA kit (Mabtech, 3485–1 H-20).
Granzyme b elisa kit
Manufactured by Mabtech
Sourced in United States, Sweden
The Granzyme B ELISA kit is a laboratory tool used to measure the concentration of Granzyme B, a cytotoxic serine protease, in biological samples. It is designed to provide accurate and reliable quantitative data on Granzyme B levels.
Automatically generated - may contain errors
Lab products found in correlation
M30 apoptosense ck18 kit, by Diapharma (1 mentions)
Human perforin elisa kit, by Abcam (1 mentions)
Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
7 aad viability staining solution, by BioLegend (1 mentions)
Human ifnγ elisa kit, by BioLegend (1 mentions)
Clariostar plus instrument, by BMG Labtech (1 mentions)
5 protocols using granzyme b elisa kit
1
Organoid-T Cell Killing Assay
2
Investigating NK Cell Cytotoxic Granules
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Granzyme B and perforin secreted from NK cells were investigated under coculture conditions with K562 tumor cells. NK cells were seeded at 40,000 cells per well and cocultured with 4,000 K562 cells per well for 3 hours. The concentrations of lytic granules were determined using a granzyme B ELISA kit (Mabtech, Cincinnati, OH, USA) and a human perforin ELISA kit (Abcam, Cambridge, MA, USA). After the experiments were performed according to the manufacturer's instructions, the results were read using a spectrophotometer (Molecular Devices), and the concentrations of granules were analyzed using GraphPad Prism 8 software.
3
Cytokine and Granzyme B Secretion in TCCs
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TCCs were stimulated with PMA and anti-CD3 (OKT3 antibody; Janssen-Ortho, Toronto, Canada) for 24 hours. Cytokine secretion of individual TCCs was measured in supernatants using a Th1/Th2/Th17 Cytokine Multi-Analyte ELISArray Kit (Qiagen) and a granzyme B ELISA kit (Mabtech, Nacka Strand, Sweden). Data were analyzed using GraphPad Prism (La Jolla, CA).
4
Evaluation of HOXB7-specific HTL Cytotoxicity
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The cytotoxic activity of HOXB7‐specific HTLs was evaluated by granzyme B ELISA and flow cytometry. Tumor cells were treated with IFN‐γ (500 IU/ml) for 48 h before coculturing with HTLs to enhance HLA‐DR expression. After coculturing HTLs with target HNSCC cells, supernatants were collected and evaluated using the Granzyme B ELISA kit (Mabtech) according to the manufacturer's instructions. In the killing assay, the target HNSCC cells were labeled using the CellTrace CFSE Cell Proliferation Kit (Invitrogen). After 6 h of coculturing with various E : T ratios, dead cells were labeled with 7‐AAD viability staining solution (BioLegend). Dead tumor cells were detected as CFSE and 7‐AAD double‐positive cells.
5
Measuring Cytokine Secretion by ELISA
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Human IFN-γ ELISA kit (Biolegend or MabTech) and granzyme B ELISA kit (MabTech) were used to measure cytokines secretion. Supernatants were collected from TICS, followed by centrifugation at 700 × g for 4 min to remove cell debris. After preparation of samples, ELISAs were conducted according to the manufacturers’ protocols. After measuring the absorbance at a wavelength of 450 nm and 570 nm, subtraction of 570 nm readings from those at 450 nm was performed on a CLARIOstar Plus instrument (BMG Labtech), followed by subtraction of an averaged background signal. IFN-γ and granzyme B concentrations were then calculated and plotted against different number of cancer cells using GraphPad software.
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