Tnfr2

Manufactured by Santa Cruz Biotechnology

Sourced in United States

TNFR2 is a cell surface receptor that binds to the cytokine tumor necrosis factor (TNF). It plays a role in regulating immune and inflammatory responses.

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Lab products found in correlation

Tnfr1, by Santa Cruz Biotechnology (3 mentions) Bca protein assay, by Thermo Fisher Scientific (2 mentions) Horseradish peroxidase conjugated secondary antibody, by GE Healthcare (1 mentions) Glial fibrillary acidic protein (gfap), by BD (1 mentions) Gap43, by Fujifilm (1 mentions) Rabbit anti β actin, by Cell Signaling Technology (1 mentions) Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Quantity one software, by Bio-Rad (1 mentions) Synaptotagmin, by Synaptic Systems (1 mentions) Tumor necrosis factor (tnf), by Santa Cruz Biotechnology (1 mentions) Mouse anti β tubulin, by Merck Group (1 mentions) Anti rat igg, by Merck Group (1 mentions) Myelin basic protein mbp, by Merck Group (1 mentions) Protease, by Roche (1 mentions) Quantity one 1 d analysis software, by Bio-Rad (1 mentions) Phosphatase inhibitor, by Roche (1 mentions) Tnf α, by Santa Cruz Biotechnology (1 mentions) X omat bt film, by Carestream (1 mentions) Mini protean 3 apparatus, by Bio-Rad (1 mentions)

5 protocols using tnfr2

Western Blot Analysis of Neural Markers

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Following blocking in 5% non-fat milk in tris buffered saline + Triton (TBS-T), membranes were probed overnight at 4°C with one of the following antibodies: recognizing glial fibrillary acidic protein (GFAP, 1:500, BD Pharmingen), growth associated protein 43 (GAP43, 1:5,000), ionized calcium binding adapter molecule 1 (Iba1, 1:400, Wako), myelin basic protein (MBP, 1:500, Millipore), toll-like receptor 4 (TLR4, 1:200, Santa Cruz), and tumor necrosis factor receptor 2 (TNFR2, 1:200, Santa Cruz). After extensive washes in TBS-T, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (1:2,000, GE Healthcare for anti-mouse and anti-rabbit, 1:1,000, Jackson ImmunoResearch for anti-rat) for 30 min at room temperature. Detection was performed with SuperSignal West Pico chemiluminescent substrate (Thermo Scientific). Quantification was performed using Quantity One software from Bio-Rad. Blots were normalized using either mouse anti-β-actin (1:500, Santa Cruz) or rabbit anti-β-actin (1:1,000, Cell Signaling).

Novrup H.G., Bracchi-Ricard V., Ellman D.G., Ricard J., Jain A., Runko E., Lyck L., Yli-Karjanmaa M., Szymkowski D.E., Pearse D.D., Lambertsen K.L, & Bethea J.R. (2014). Central but not systemic administration of XPro1595 is therapeutic following moderate spinal cord injury in mice. Journal of Neuroinflammation, 11, 159.

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Protein expression analysis of mouse hippocampus

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Mice were transcardially perfused with cold 0.1 M PBS pH 7.4, the hippocampi dissected out and homogenized in RIPA buffer (0.01 M Sodium Phosphate pH 7.2, 0.15 M NaCl, 1% Nonidet-40, 1% Sodium Deoxycholate, 0.1% SDS, 2 mM EDTA, containing protease (Roche Diagnostics, Germany) and phosphatase inhibitors (Sigma, USA). Protein samples were electrophoresed onto 6–15% sodium dodecyl sulfate-polyacrylamide gels and transferred to nitrocellulose membranes, which were blocked with 5% milk and incubated with antibodies against the following proteins: growth associated protein 43 (GAP43) 1:1000 (Genetex), microtubule-associated protein 2 (MAP2) 1:800, myelin basic protein (MBP) 1:1000 (Millipore), myelin proteolipid protein (PLP) 1:400 (Pierce Biotechnology), synaptotagmin 1:1000 (Synaptic Systems), TNF 1:100, TNFR1 1:300 and TNFR2 1:300 (Santa Cruz). Peroxidase-conjugated anti-rabbit, or anti-mouse IgG (1:5,000, Bio-Rad), or anti-rat IgG (1:2,000, Sigma) were used as secondary antibody. Signal was detected by SuperSignal WestPico Chemiluminescent Substrate (Pierce) according to the manufacturer’s instructions. Membranes were reprobed with mouse anti-β-tubulin (1:10,000, 1 h, RT, Sigma). Quantification of western blots was performed by densitometry using Quantity One, 1-D Analysis software (Bio-Rad), normalizing each band to the β-tubulin signal.

Anna D., Paul M., Roberta B., Winston W., Spencer S., Danielle B., Mariagrazia G, & R B.J. (2014). NEUROPATHIC PAIN-INDUCED DEPRESSIVE-LIKE BEHAVIOR AND HIPPOCAMPAL NEUROGENESIS AND PLASTICITY ARE DEPENDENT ON TNFR1 SIGNALING. Brain, behavior, and immunity, 41, 65-81.

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Western Blot Analysis of Inflammatory Proteins

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To obtain the protein, tissue samples and cells were lysed using RIPA lysis buffer and centrifuged at 12,000×g for 20 min to collect the supernatant without lipid. Protein concentration was measured using the BCA protein assay (Thermo Scientific, MA, USA). Equal amounts of protein were loaded onto an SDS-polyacrylamide gel, electrophoresed, and transferred to polyvinylidene difluoride membranes (PVDF, Minipore, MA, USA). The membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) at room temperature for 1 h, and then incubated with primary antibodies at 4°C for overnight. Each membrane was washed with TBST three times for 15 min followed by incubation with an HRP-conjugated secondary antibody (Zhongshan Jiangqiao, Beijing, China) at room temperature for 1 h. Finally, each membrane was developed using an enhanced chemiluminescence (ECL) detection kit (Minipore, MA, USA) and visualized using X-OMAT BT film (Carestream, Toronto, Canada). The antibodies for TNF-α, TNFR1, and TNFR2 were purchased from Santa Cruz (California, USA), the primary antibodies for AR, IκB, NF-κB, and P65 from Proteintech Group (Chicago, USA), TNF-α from CST (Boston, USA), and β-actin, and GAPDH from Zenbio Biotech Co., Ltd. (Chengdu, China).

Lang Q., Yidong X., Xueguang Z., Sixian W., Wenming X, & Tao Z. (2019). ETA-mediated anti-TNF-α therapy ameliorates the phenotype of PCOS model induced by letrozole. PLoS ONE, 14(6), e0217495.

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Immunoblotting Analysis of Neuroreceptor Proteins

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Western blotting assay was prepared as previously described [14 (link)]. A 10% polyacrylamide gel and Bio-Rad mini-Protean III apparatus (Bio-Rad) were used to separate the membrane-enriched proteins (15 µg) by electrophoresis (SDS-PAGE; 90 V). The proteins were blotted onto a nitrocellulose membrane (Bio-Rad) and incubated overnight with the specific primary antibodies (1:1000) TNFR2 (sc-7862, Santa Cruz Biotechnology), NR2A, pAMPA, AMPA, and NR1 (4205, 8084, 13,185, and 5704, Cell Signaling Technology). The membranes were then incubated for 2 h with a specific secondary antibody.
Proteins recognized by antibodies were detected using electrochemiluminescence (ECL). To standardize and quantify the immunoblots, we used ImageJ software (NIH). The β-ACTIN antibody (sc-1616; Santa Cruz Biotechnology) was used as an internal experimental control, with results expressed concerning β-ACTIN intensity.

Kinoshita P.F., Orellana A.M., Andreotti D.Z., de Souza G.A., de Mello N.P., de Sá Lima L., Kawamoto E.M, & Scavone C. (2022). Consequences of the Lack of TNFR1 in Ouabain Response in the Hippocampus of C57BL/6J Mice. Biomedicines, 10(11), 2937.

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Quantification and Western Blot Analysis

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Total protein from cell cultures and cervical tissues was quantified by the Bradford assay. Western blot assay was performed as described [48 (link)] with the primary antibody for PGRN (1:1000, Abcam), phospho-Akt (Thr308, 1:1000), phospho-Akt (Ser473, 1:2000), total Akt (1:1000), phospho-Erk1/2 (Thr202/Tyr204, 1:2000) and total Erk1/2 (1:1000), phospho-TSC-2 (Thr1462, 1:1000), total TSC-2 (1:1000), phospho-mTOR-Ser2448 (1:1000), phospho-mTOR-Ser2481 (1:1000), total mTOR (1:1000), phospho-p70S6K (Thr389, 1:1000), total p70S6K (1:1000), phospho-4E-BP1 (Thr37/46, 1:1000), total 4E-BP1 (1:1000), phospho-PKCα (Thr638, 1:1000), total PKCα (1:1000, all Cell Signaling Technology), TNFR1 (1:1000), TNFR2 (1:1000, Santa Cruz Biotechnology, Dallas, TX), c-myc (1:2000) and cyclin D1 (1:2000, both ProteinTech Group, Chicago, IL). GAPDH antibody was a control (1:2000, Hangzhou Goodhere Biotech, China).

Feng T., Zheng L., Liu F., Xu X., Mao S., Wang X., Liu J., Lu Y., Zhao W., Yu X, & Tang W. (2016). Growth factor progranulin promotes tumorigenesis of cervical cancer via PI3K/Akt/mTOR signaling pathway. Oncotarget, 7(36), 58381-58395.

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