To analyse spontaneous and inducible CXCL16 secretion by tumour cells 104 Panc02-OVA or 2 x 104 T110299-OVA tumour cells were seeded into a 96-well flat bottom plate and stimulated with 20 ng/ml IFN-γ (Peprotech), 20 ng/ml TNF-γ (Peprotech) or a combination of both for 48 h. To analyse intracellular and transmembrane CXCL16 concentration 2 x 105 Panc02-OVA or 2 x 105 T110299-OVA were plated in a 6-well plate and stimulated with or without 20 ng/ml IFN-γ. After 72 h supernatants were harvested, cells were washed once with PBS and cells were lysed using RIPA Lysis Buffer system (Santa Cruz Biotechnology). CXCL16 concentrations in supernatants and lysates were determined using ELISA (R&D systems). Absorbance was measured with Mithras LB 940 Multimode Microplate reader (Software MicroWin 2000). To quantify CXCL16 secretion by human pancreatic tumour cell lines, 2 x 105 cells were seeded in a 6-well plate and supernatants were analysed after 72 h by ELISA (R&D systems).
Tnf γ
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TNF-γ is a lab equipment product designed to detect and measure the presence of tumor necrosis factor-gamma (TNF-γ) in biological samples. It provides a reliable and efficient method for researchers to quantify this important cytokine, which plays a crucial role in immune system regulation and inflammatory processes.
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Lab products found in correlation
3 protocols using tnf γ
1
CXCL16 Secretion by Tumor Cells
2
Regulation of Corneal Keratocyte Apoptosis
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A mouse corneal keratocyte cell line was transfected with agomir-122 (20 μM), antagomir-122 (20 μM), or a plasmid carrying CPEB1 cDNA. Alternatively, cells were transfected with a plasmid carrying CPEB1 cDNA with wild-type or miR-122-binding site-mutated 3′-UTR. The cells were then treated with or without a mixed cytokine cocktail containing 100 ng/ml TNF-α, IL-1β and TNF-γ (Peprotech, Rocky Hill, NJ, USA). After 48 h, the cells were stained with annexin V (Keygen Biotech, Nanjing, China) per the manufacturer’s instructions, and the percentage of apoptotic cells (annexin V+) was determined by flow cytometry.
3
CXCL16 Secretion by Tumor Cells
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To analyse spontaneous and inducible CXCL16 secretion by tumour cells 104 Panc02-OVA or 2 x 104 T110299-OVA tumour cells were seeded into a 96-well flat bottom plate and stimulated with 20 ng/ml IFN-γ (Peprotech), 20 ng/ml TNF-γ (Peprotech) or a combination of both for 48 h. To analyse intracellular and transmembrane CXCL16 concentration 2 x 105 Panc02-OVA or 2 x 105 T110299-OVA were plated in a 6-well plate and stimulated with or without 20 ng/ml IFN-γ. After 72 h supernatants were harvested, cells were washed once with PBS and cells were lysed using RIPA Lysis Buffer system (Santa Cruz Biotechnology). CXCL16 concentrations in supernatants and lysates were determined using ELISA (R&D systems). Absorbance was measured with Mithras LB 940 Multimode Microplate reader (Software MicroWin 2000). To quantify CXCL16 secretion by human pancreatic tumour cell lines, 2 x 105 cells were seeded in a 6-well plate and supernatants were analysed after 72 h by ELISA (R&D systems).
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