HNSCC cells (2 × 105) were seeded in 24-well plates overnight. The next day, PBMCs (3 × 106) were cocultured with the HNSCC cells with/without 10 µg/mL CDA for 2 and 6 hours. For transwell cocultures, 2 × 105 HNSCC cells were seeded into the bottom compartment of a 24-well plate overnight. PBMCs (3 × 106) were seeded into the top 0.4-µm ThinCert cell culture insert (Greiner Bio-One, catalog no. 665641) the next day and cultured for 6 hours ± 10 µg/mL CDA. Supernatants were harvested and the levels of IFNβ, IL6, TNFα, and IL1β were assessed with human type-I IFN (Biorbyt, catalog no. orb561974), TNFα (Invitrogen, catalog no. 88–7346), IL6 (Invitrogen, catalog no. 88–7066), and IL1β (Invitrogen, catalog no. 88–7261) ELISA kits following the manufacturer's instructions.
Tnf α
Manufactured by Biorbyt
Sourced in United Kingdom
TNF-α is a protein that is involved in the regulation of various cellular processes, including inflammation, immune response, and cell death. It plays a crucial role in the body's defense against infection and disease. This product is a laboratory reagent used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Il 1β, by Thermo Fisher Scientific (2 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Il 1β, by Biorbyt (1 mentions)
Interleukin 6 (il 6), by Biorbyt (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Thincert cell culture insert, by Greiner (1 mentions)
Il 17a, by Thermo Fisher Scientific (1 mentions)
Prism 6, by GraphPad (1 mentions)
Spss 19, by IBM (1 mentions)
Caspase 3, by Santa Cruz Biotechnology (1 mentions)
Anti nf κb, by Santa Cruz Biotechnology (1 mentions)
3 protocols using tnf α
1
HNSCC-PBMC Coculture Cytokine Analysis
2
Resveratrol Modulates Inflammatory Cytokines in Mice with Depressive Disorder
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The serum levels of IL-8 (cat. no. KHC0081; Thermo Fisher Scientific, Inc.), IL-1β (cat. no. MBS700340; Thermo Fisher Scientific, Inc.), IL-17A (cat. no. 88-7876-88; Thermo Fisher Scientific, Inc.), H3R (cat no. orb154346; Biorbyt, Cambridge, UK) and TNF-α (cat. no. KHC3014; Thermo Fisher Scientific, Inc.), were detected in mice with depressive disorder prior to and following treatment with resveratrol using ELISA kits according to the manufacturer's protocol. Finally, the serum concentration levels of IL-8, IL-1β, IL-17A and TNF-α were measured using an enzyme microplate reader at 450 nm.
Statistical analysis. The results are presented as the mean ± standard error of the mean. Significance was established with SPSS 19.0 (IBM Corp., Armonk, NY, USA) and GraphPad Prism 6 software (GraphPad Software, Inc., La Jolla, CA, USA). A Kaplan-Meier plot, log-rank test, multi-variant Cox regression analysis, Pearson's correlation coefficient and a two-tailed Student's t-test were used. P<0.05 was considered to indicate a statistically significant difference.
Statistical analysis. The results are presented as the mean ± standard error of the mean. Significance was established with SPSS 19.0 (IBM Corp., Armonk, NY, USA) and GraphPad Prism 6 software (GraphPad Software, Inc., La Jolla, CA, USA). A Kaplan-Meier plot, log-rank test, multi-variant Cox regression analysis, Pearson's correlation coefficient and a two-tailed Student's t-test were used. P<0.05 was considered to indicate a statistically significant difference.
3
Immunohistochemical Analysis of Apoptosis Markers
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Immunohistochemistry of coronal sections was based on the EnVision antibody method using a speci c antibody Caspase-3, anti-NF-κB (1:100 dilution; Santa Cruz Biotechnology) and TNF-α (orb338920; Biorbyt, England). Brie y, the samples were washed in three stages with phosphate buffered saline (PBS). The secondary antibody was added to the sample at a dilution of 1:150, and then the samples were incubated at 37°C for 1 h and 30 min. The sample was then transferred from the incubator to the dark room and after 4 times washing, DAPI was added followed by PBS after 5 min [25] . Eventually the samples were viewed by a Labomed tcs400 uorescent microscope, photographs were taken from each slide and analyzed using ImageJ software (version 1.49, NIH).
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