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Mtnf α

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MTNF-α is a recombinant human Tumor Necrosis Factor-alpha (TNF-α) protein. TNF-α is a cytokine that plays a central role in inflammation and immune response.

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Lab products found in correlation

Mifn γ, by BD (3 mentions) Mtgf β, by R&D Systems (1 mentions) Proteome profiler array mouse cytokine array panel a, by R&D Systems (1 mentions) Mmcp 1, by R&D Systems (1 mentions) Live dead fixable green dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm, by BD (1 mentions) Cytoflex s flow cytometer, by Beckman Coulter (1 mentions)

4 protocols using mtnf α

1

Quantifying Cytokine Levels in Lung Samples

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mTGF-β (R&D system, Minneapolis, MN), mIL-17A (biolegend, San Diego, CA), mTNF-α (BD biosciences, San Jose, CA, USA), mIFN-γ (BD biosciences), mIL-1β (BD biosciences) concentrations in BAL fluid or in lung homogenates were measured by ELISA according to the manufacturer's instructions.
Monnier J, & Zabel B.A. (2014). Anti-Asialo GM1 NK Cell Depleting Antibody Does Not Alter the Development of Bleomycin Induced Pulmonary Fibrosis. PLoS ONE, 9(6), e99350.
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2

Cytokine and ALT Measurement

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Sera were assayed for cytokine production by ELISA according to the manufacturers' instructions. The kits utilized were: mIL-10, mIFNγ, mTNFα (BD-Pharmingen, La Jolla, CA). Alanine aminotransferase (ALT) sera levels were determined using a specific colorimetric kit (Wiener Laboratories) and by following the manufacturer's instructions.
Barrios B., Baez N.S., Reynolds D., Iribarren P., Cejas H., Young H.A, & Rodriguez-Galan M.C. (2014). Abrogation of TNFα Production during Cancer Immunotherapy Is Crucial for Suppressing Side Effects Due to the Systemic Expression of IL-12. PLoS ONE, 9(2), e90116.
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3

Cytokine Profiling in Infected Cells

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Proteome profiler array (mouse cytokine array panel A, R&D Biosystems) was used to assay the level of 40 cytokines according to the manufacturer’s instructions. Briefly, the capture antibody of cytokines and chemokines were spotted on nitrocellulose membranes. The sample/antibody mixture was incubated and the cytokine-chemokine antibody complex bound to the immobilized capture antibody on the membrane. After adding streptavidin-horseradish peroxidase and chemiluminescent reagents, the signal is proportional to the amount of cytokine bound. For secreted cytokines, supernatants were collected from infected/treated cells at the indicated times. mTNF-α (BD Pharmingen) and mMCP-1 (R&D Biosystems) was measured using an ELISA kit according to the manufacturer’s instructions.
Das S., Sarkar A., Choudhury S.S., Owen K.A., Derr-Castillo V.L., Fox S., Eckmann L., Elliott M.R., Casanova J.E, & Ernst P.B. (2015). Engulfment and Cell Motility Protein 1 (ELMO1) Has an Essential Role in the Internalization of Salmonella Typhimurium Into Enteric Macrophages That Impact Disease Outcome. Cellular and Molecular Gastroenterology and Hepatology, 1(3), 311-324.
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4

Staining and Quantification of Immune Cells

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For the staining of surface proteins, cells were resuspended in FACS buffer (0.1% BSA in PBS) with the respective antibodies and incubated for 30 minutes on ice and in the dark. The following antibodies were used: mCD4 (eBioscience), mCD8 (BD Biosciences), mCD45.1 (eBioscience), mCD45.2, mIFN-γ, mTNF-α (all from BD Biosciences); hCD4 (Thermo Fisher Scientific), hCD8 (Dako). Viability of cells was determined using a live/dead cell marker (LIVE/DEAD Fixable Green Dead Cell Stain Kit, Thermo Fisher Scientific). For intracellular cytokine staining, cells were permeabilized by resuspending them in Cytofix/Cytoperm (BD Biosciences) prior to the antibody staining, following the manufacturer’s instructions. In order to determine the absolute cell count by flow cytometry, CountBright™ Absolute Counting Beads (Thermo Fisher Scientific) were added to the cell suspension shortly before measurement. If CAR expression was analyzed together with other surface proteins such as CD8, stainings were performed sequentially. The CAR was first stained with an anti-human IgG antibody (Abcam) followed by the staining of the other surface proteins with the respective antibody. The samples were then analyzed on a CytoFLEX S flow cytometer (Beckman Coulter). The obtained data was evaluated with FlowJo 10.4.
Klopp A., Schreiber S., Kosinska A.D., Pulé M., Protzer U, & Wisskirchen K. (2021). Depletion of T cells via Inducible Caspase 9 Increases Safety of Adoptive T-Cell Therapy Against Chronic Hepatitis B. Frontiers in Immunology, 12, 734246.
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