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Recombinant human tnf α protein

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Recombinant human TNF-α protein is a soluble cytokine produced using recombinant DNA technology. It is a key regulator of immune responses and inflammatory processes.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Recombinant human tgf β1, by Thermo Fisher Scientific (2 mentions) Gyy4137, by Cayman Chemical (1 mentions) Accumax, by Merck Group (1 mentions) Anti arg1, by Cell Signaling Technology (1 mentions) Anti bax, by Cell Signaling Technology (1 mentions) Anti bcl 2, by Cell Signaling Technology (1 mentions) Anti p65, by Cell Signaling Technology (1 mentions) Anti p p65, by Cell Signaling Technology (1 mentions) Anti ho 1, by Cell Signaling Technology (1 mentions) Anti traf1, by Cell Signaling Technology (1 mentions) Anti il 10, by Cell Signaling Technology (1 mentions) Anti inos, by Cell Signaling Technology (1 mentions) Rhodamine b, by Merck Group (1 mentions) Anti cox 2, by Cell Signaling Technology (1 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Clodronate liposome, by Liposoma (1 mentions) Anti phospho p38 mapk thr180 tyr182, by Cell Signaling Technology (1 mentions) Ecl western blotting detection system, by PerkinElmer (1 mentions)

Close spelling variants of this product

TNF-α) recombinant protein Recombinant human TNF-α Recombinant TNF-α TNF α) protein Recombinant human TNF Human TNF-α TNF-α Recombinant proteins

11 protocols using recombinant human tnf α protein

1

Inducing RPE-EMT and EMT in Cell Lines

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For inducing TGF–β signaling-associated RPE–EMT in hRPE monolayers, cells were co-treated with 20 ng/mL of recombinant human TGF–β1 (Thermo Scientific, Waltham, MA, USA; Catalog # PHG9204) and recombinant human TNF–α protein (R&D systems, Minneapolis, MN, USA Catalog # 210-TA-020) in RPE medium for 24 h in 37 °C/5% CO2 conditions. For inducing EMT in A549 and H1299 lines, cells were treated with 20 ng/mL of recombinant human TGF–β1 only.
Sripathi S.R., Hu M.W., Turaga R.C., Mikeasky R., Satyanarayana G., Cheng J., Duan Y., Maruotti J., Wahlin K.J., Berlinicke C.A., Qian J., Esumi N, & Zack D.J. (2023). IKKβ Inhibition Attenuates Epithelial Mesenchymal Transition of Human Stem Cell-Derived Retinal Pigment Epithelium. Cells, 12(8), 1155.
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2

Recombinant human TNF-α Protein Assay

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Recombinant human TNF-α protein was purchased from R&D systems (Minneapolis, MN, USA). The slow-releasing H2S donor molecule, GYY4137, was purchased from Cayman (Ann Arbor, MI, USA). Bovine Serum Albumin (BSA) was purchased from Millipore Sigma (Burlington, MA, USA).
Escaffre O., Szaniszlo P., Törő G., Vilas C.L., Servantes B.J., Lopez E., Juelich T.L., Levine C.B., McLellan S.L., Cardenas J.C., Freiberg A.N, & Módis K. (2023). Hydrogen Sulfide Ameliorates SARS-CoV-2-Associated Lung Endothelial Barrier Disruption. Biomedicines, 11(7), 1790.
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3

Inducing RPE-EMT via TGF-β and TNF-α

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The hRPE monolayers were incubated in proteo-collagenolytic enzyme (AccuMAX, Sigma-Aldrich, St. Louis, MO) for 15 to 20 minutes. Gentle mechanical trituration was performed by pipetting approximately 15 times with a P1000 pipette. AccuMAX was neutralized by adding double the amount of RPE medium and AccuMAX was removed by aspiration after cells were centrifuged at 150×g for 5 minutes. Cell viability was assessed by Trypan blue dye exclusion. Cells were then resuspended in RPE medium and plated on Matrigel-coated plates at a density of 30,000 cells/cm2 and incubate at 37°C/5% CO2. For inducing TGF-β signaling associated RPE-EMT, the hRPE monolayers were cultured for approximately 3 months before cotreatment with equal concentration of 1 to 40 ng/mL of recombinant human TGF-β1 (Thermo Fisher Scientific, Waltham, MA; Catalog # PHG9204) and recombinant human TNF-α protein (R&D Systems. Minneapolis, MN; Catalog # 210-TA-020) in RPE medium for 24, 48, and 72 hours in 37°C/5% CO2.
Sripathi S.R., Hu M.W., Liu M.M., Wan J., Cheng J., Duan Y., Mertz J.L., Wahlin K.J., Maruotti J., Berlinicke C.A., Qian J, & Zack D.J. (2021). Transcriptome Landscape of Epithelial to Mesenchymal Transition of Human Stem Cell–Derived RPE. Investigative Ophthalmology & Visual Science, 62(4), 1.
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4

Macrophage Polarization Cytokine Assay

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Recombinant human TNF-α protein was purchased from R&D (Boston, MO, USA). Clodronate liposome was purchased from Liposoma B.V (Vrije University, Netherlands). Rhodamine B and LPS were purchased from Sigma (St. Louis, MO, USA). Anti-Arg1, anti-IL-10, anti-iNOS, anti-TRAF1, anti-p-p65, anti-p65, anti-Cox2, anti-Ho-1, anti-Bcl2, anti-Bax, and anti-GAPDH antibodies were purchased from Cell Signaling Technology (Boston, MO, USA). SPIONs were provided by Professor Ning Gu’s team at Southeast University, Nanjing, PR China.
Xu Y., Liu X., Li Y., Dou H., Liang H, & Hou Y. (2021). SPION-MSCs enhance therapeutic efficacy in sepsis by regulating MSC-expressed TRAF1-dependent macrophage polarization. Stem Cell Research & Therapy, 12, 531.
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5

Signaling Pathway Analysis Protocol

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Dulbecco’s modified Eagle’s medium (DMEM)/F-12 and fetal bovine serum were purchased from Thermo Fisher Scientific (Grand Island, NY, USA). BioTraceTM NT membrane was purchased from Pall Life Sciences (Ann Arbor, MI, USA). The enhanced chemiluminescence (ECL) Western blotting detection system was purchased from Perkin Elmer (Waltham, MA, USA). Anti-phospho-EGFR (Tyr1173, Cat#4407), anti-phospho-p38 MAPK (Thr180/Tyr182, Cat#9211), anti-phospho-JNK1/2 (Thr183/Tyr185, Cat#9255), anti-phospho-FoxO1 (Ser256, Cat#9461), and anti-phospho-p65 (Ser536, Cat#3033) antibodies were purchased from Cell Signaling (Danvers, MA, USA). Antibodies of anti-TNFR1 (Cat#sc-52739), anti-TNFR2 (Cat#sc-8041), and p38 MAPK inhibitor (p38i) VIII were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-GAPDH (Cat#MCA-1D4) was purchased from EnCor Biotechnology (Gainesville, FL, USA). AG1478, SP600125, and Tanshinone IIA were purchased from Enzo Life Science (Farmingdale, NY, USA). AS1842856 was obtained from EMD Millipore (Billerica, MA, USA). Helenalin was purchased from Cayman Chemical (Ann Arbor, MI, USA). Recombinant human TNF-α protein was purchased from R&D Systems (Minneapolis, MN, USA). TRIzol reagent, enzymes, and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Yang C.M., Yang C.C., Hsu W.H., Hsiao L.D., Tseng H.C, & Shih Y.F. (2022). Tumor Necrosis Factor-α-Induced C-C Motif Chemokine Ligand 20 Expression through TNF Receptor 1-Dependent Activation of EGFR/p38 MAPK and JNK1/2/FoxO1 or the NF-κB Pathway in Human Cardiac Fibroblasts. International Journal of Molecular Sciences, 23(16), 9086.
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6

Fibroblast Culture and TNFα Treatment

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Fibroblasts were obtained from skin biopsies of four patients (age at biopsy range: 8 to 19 years) and six controls (age at biopsy range: 17 years to adult). Corresponding informed consent and institutional ethics approval were obtained (Ethics Committee of the Universitat de Barcelona, IRB00003099), and all methods were performed in accordance with the relevant guidelines and regulations. Fibroblasts were cultured in DMEM supplemented with 10% FBS (Gibco, LifeTechnologies) and 1% penicillin-streptomycin (Gibco, LifeTechnologies) and were maintained at 37ºC and 5% CO2. When appropriate, cells were treated with 10 ng/μl recombinant human TNFα protein (R&D systems) for 6, 24 or 48 hours.
Castilla-Vallmanya L., Selmer K.K., Dimartino C., Rabionet R., Blanco-Sánchez B., Yang S., Reijnders M.R., van Essen A.J., Oufadem M., Vigeland M.D., Stadheim B., Houge G., Cox H., Kingston H., Clayton-Smith J., Innis J.W., Iascone M., Cereda A., Gabbiadini S., Chung W.K., Sanders V., Charrow J., Bryant E., Millichap J., Vitobello A., Thauvin C., Mau-Them F.T., Faivre L., Lesca G., Labalme A., Rougeot C., Chatron N., Sanlaville D., Christensen K.M., Kirby A., Lewandowski R., Gannaway R., Aly M., Lehman A., Clarke L., Graul-Neumann L., Zweier C., Lessel D., Lozic B., Aukrust I., Peretz R., Stratton R., Smol T., Dieux-Coëslier A., Meira J., Wohler E., Sobreira N., Beaver E.M., Heeley J., Briere L.C., High F.A., Sweetser D.A., Walker M.A., Keegan C.E., Jayakar P., Shinawi M., Kerstjens-Frederikse W.S., Earl D.L., Siu V.M., Reesor E., Yao T., Hegele R.A., Vaske O.M., Rego S., Shapiro K.A., Wong B., Gambello M.J., McDonald M., Karlowicz D., Colombo R., Serretti A., Pais L., O’Donnell-Luria A., Wray A., Sadedin S., Chong B., Tan T.Y., Christodoulou J., White S.M., Slavotinek A., Barbouth D., Swols D.M., Parisot M., Bole-Feysot C., Nitschké P., Pingault V., Munnich A., Cho M.T., Cormier-Daire V., Balcells S., Lyonnet S., Grinberg D., Amiel J., Urreizti R, & Gordon C.T. (2020). Phenotypic spectrum and transcriptomic profile associated with germline variants in TRAF7. Genetics in medicine : official journal of the American College of Medical Genetics, 22(7), 1215-1226.
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7

Hypoxic and Inflammatory Modulation of IPTD-MSCs

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Recombinant human TNF-α protein (R&D Systems, Minneapolis, MN) was reconstituted in research-grade water (Hospira, Lake Forest, IL) to a concentration of 100 ng/mL, aliquoted, and stored at − 20 °C. Dimethyloxallyl glycine (DMOG; Cayman Chemicals, Ann Arbor, MI) was dissolved in water to yield a stock solution of 57.1 mM, aliquoted (100 μL), and stored at − 80 °C. IPTD-MSCs were incubated in T-25 flasks in 5 ml of CMRL-1066 medium supplemented with 5% hPL until ~ 50% confluent. Cells were cultured for 24 h in CMRL 1066 medium, or medium containing 10 ng/ml TNF-α, 1 mM DMOG or 1 mM DMOG and 10 ng/ml TNF-α. Following treatment, cells were collected in 1.7-ml Eppendorf tubes and stored in RLT buffer (Qiagen, Germantown, MD) at − 80 °C for future preparation of cDNA.
Khiatah B., Qi M., Du W., T-Chen K., van Megen K.M., Perez R.G., Isenberg J.S., Kandeel F., Roep B.O., Ku H.T, & Al-Abdullah I.H. (2019). Intra-pancreatic tissue-derived mesenchymal stromal cells: a promising therapeutic potential with anti-inflammatory and pro-angiogenic profiles. Stem Cell Research & Therapy, 10, 322.
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8

Development and Characterization of Infliximab-Loaded Lipid Carriers

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Infliximab (European Pharmacopeia reference standard) and coconut oil (from Cocos nucifera) were purchased from Merck Life Sciences, UK. Gelucire® 48/16 (also known as Polyethylene glycol-32 (PEG-32) stearate; and is composed of PEG-32 (molecular weight (MW) 1500 g/mol) esters of palmitic (C16) and stearic (C18) acids) and 44/14 (also known as lauroyl PEG-32 glycerides; and is composed of PEG-32 (MW 1500 g/mol) mono- and diesters of lauric acid (C12) mixed with a small portion of mono, di- and triglycerides) were kindly donated by Gattefossé, France. The recombinant human TNF-α protein was acquired from R&D Systems, Bio-Techne, UK. The goat anti-human IgG Fc secondary antibody Horseradish Peroxidase (HRP) conjugate was purchased from ThermoFisher Scientific, UK. Bovine Serum Albumin (BSA), Tween 20 and Dulbecco's Phosphate Buffered Saline (PBS) were obtained from Sigma-Aldrich, UK. The enhanced K-BLUE 3,3',5,5' tetramethylbenzidine (TMB) substrate and RED STOP solutions were bought from Neogen Europe Limited, Scotland. Ultrapure water was acquired by filtering tap water through a PURELAB Chorus 1 complete water purification system (Veolia Water Technologies, UK).
Awad A., Goyanes A., Orlu M., Gaisford S, & Basit A.W. (2023). 3D printed infliximab suppositories for rectal biologic delivery. International Journal of Pharmaceutics: X, 5, 100176.
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9

NF-κB Luciferase Reporter Assay

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NF-κB Luciferase Reporter Vector was purchased from Signosis (Santa Clara, CA, USA) and transfected into HEK-293 cells using FuGENE HD (Promega, Fitchburg, MA, USA) following the manufacturer’s protocol. The transfected cells were incubated in serum-free DMEM for 16 h and then treated with DMSO or indicated compounds. After 3 h, cells were treated with or without 50 ng/mL of Recombinant Human TNFα Protein (R&D systems, Minneapolis, MN, USA) for another 3 h. The protein lysates were analyzed using Steady-Glo® Luciferase Assay System (Promega, Fitchburg, MA, USA) and TECAN Infinite 200 pro (Männedorf, Switzerland).
Darwish S.S., Chen P.J., Hamed M.M., Wagdy R.A., Chen S.H., Abadi A.H., Abdel-Halim M., Hwang T.L, & Engel M. (2022). Development of (4-Phenylamino)quinazoline Alkylthiourea Derivatives as Novel NF-κB Inhibitors. Pharmaceuticals, 15(7), 778.
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10

Recombinant Cytokine Uptake Assay

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Recombinant human TNF-α protein was purchased from R&D System (Minneapolis, MN). FITC-conjugated dextran (average mol wt 70,000) and non-FITC-conjugated dextran (average mol wt 64,000-76,000) were purchased from SIGMA-ALDRICH (St. Louis, MO).
Miyazaki K., Hashimoto K., Sato M., Watanabe M., Tomikawa N., Kanno S., Kawasaki Y., Momoi N, & Hosoya M. (2017). Establishment of a method for evaluating endothelial cell injury by TNF-α in vitro for clarifying the pathophysiology of virus-associated acute encephalopathy. Pediatric research, 81(6).
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