Rabbit anti tnf α

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-TNF-α is a primary antibody that recognizes human tumor necrosis factor alpha (TNF-α). It is used in various immunological techniques, such as Western blotting and ELISA, to detect and quantify TNF-α levels in biological samples.

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Lab products found in correlation

Rabbit anti il 1β, by Cell Signaling Technology (2 mentions) Rabbit anti nf κb p65, by Cell Signaling Technology (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Mouse anti lamin a c, by Active Motif (1 mentions) Rabbit anti lamin b1, by Abcam (1 mentions) Mouse anti β actin, by Merck Group (1 mentions) Laemmli sample buffer, by Bio-Rad (1 mentions) Rabbit anti il 10, by Cell Signaling Technology (1 mentions) Ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Gel imaging system, by Thermo Fisher Scientific (1 mentions) Ecl plus kit, by Thermo Fisher Scientific (1 mentions) Mouse anti stat3, by Cell Signaling Technology (1 mentions) Mouse anti perk1 2, by Cell Signaling Technology (1 mentions) Ripa lysis buffer, by Merck Group (1 mentions) Mouse anti erk1 2, by Cell Signaling Technology (1 mentions) Mouse anti β actin, by Abcam (1 mentions) Mouse anti pstat3, by Cell Signaling Technology (1 mentions) Emz104, by Kerafast (1 mentions) Aperio imagescope software, by Leica (1 mentions)

Close spelling variants of this product

TNF-α (257 citations) Anti-TNF-α (48 citations)

9 protocols using rabbit anti tnf α

Western Blot Analysis of Inflammatory Markers

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Whole‐cell lysates were generated using Laemmli sample buffer (Bio‐Rad) and diluted in SDS‐PAGE sample buffer. Cell lysates were separated in 10% or 15% SDS‐PAGE and then transferred onto nitrocellulose membranes. The membranes were blocked with 5% milk and probed with the following antibodies: rabbit anti‐lamin‐B1 (1:5,000; Abcam), mouse anti‐lamin‐A/C (1:5,000; Active Motif), rabbit anti‐TNF‐α (1:1,000; Cell Signaling), rabbit anti‐IL‐1β (1:1,000; Cell Signaling), rabbit anti‐IL‐6 (1:1,000; Novus biological), rabbit anti‐IL‐1α (1:2000, Santa Cruz), and mouse anti‐β‐actin (1:4,000; Sigma, AC‐15). Antibodies were detected with HRP‐conjugated anti‐mouse (1:10,000) or anti‐rabbit (1:10,000) antibodies and West Pico Substrate (Thermo Scientific).

Yue S., Zheng X, & Zheng Y. (2019). Cell‐type‐specific role of lamin‐B1 in thymus development and its inflammation‐driven reduction in thymus aging. Aging Cell, 18(4), e12952.

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Western Blot Analysis of Apoptosis Markers

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The penumbra area was separated from the ischemic hemisphere and homogenized as reported previously.(11 (link)) The protein concentration was determined by the Pierce™ BCA Protein Assay Kit (Thermo Scientific, IL). 50–100 µg of protein samples of each group was loaded onto 10% polyacrylamide gel, electrophoresed, and transferred to a nitrocellulose membrane. The nitrocellulose membranes were blocked and incubated with the primary antibody of rabbit anti cleaved caspase-3 (#9661), rabbit anti TNF-α (#3707) and rabbit anti IL-10 (#12163) overnight at 4 °C (Cell Signaling Technology, Inc.). The nitrocellulose membranes were then processed with secondary antibodies (Santa Cruz, Inc., USA) for 1 h at room temperature. Immuno-blots were detected using ECL Western Blotting Substrate (Thermo Fisher Scientific Inc., IL) and the membrane was detected by Carestream Molecular Imaging Software (Standard Edition).

Chen C., Xi C., Liang X., Ma J., Su D., Abel T, & Liu R. (2016). The role of kappa opioid receptor in brain ischemia. Critical care medicine, 44(12), e1219-e1225.

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Western Blotting of Inflammatory Markers

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Total protein was exacted using RIPA lysis buffer (Sigma) and protease/phosphatase inhibitors. 50 μg of protein from each sample was separated by 12% SDS-PAGE and then transferred onto a PVDF membrane (Millipore, Billerica, MA, USA). The blots were blocked with TBST containing 5% non-fat milk and incubated with primary antibodies as follows: rabbit anti-TNFα (Cell Signaling Technology, 1 : 500), rabbit anti-IL-1β (Cell Signaling Technology, 1 : 500), rabbit anti-FGF1 (Cell Signaling Technology, 1 : 400), mouse anti-ERK1/2 (Cell Signaling Technology, 1 : 400), mouse anti-pERK1/2 (Cell Signaling Technology, 1 : 200), mouse anti-STAT3 (Cell Signaling Technology, 1 : 500), mouse anti-pSTAT3 (Cell Signaling Technology, 1 : 200) and mouse anti-β-actin (Abcam, 1 : 1000) overnight at 4 °C. After rinsing with TBST, membranes were incubated with horseradish peroxidase-linked anti-rabbit or anti-mouse IgG (Santa Cruz Biotechnology, 1 : 2000) for 1 hour at room temperature. Membranes were processed with ECL Plus kit (Thermo Fisher Scientific) and exposed in a Gel imaging system (Thermo Fisher Scientific). Relative intensities of the proteins were normalized to β-actin.

Zhang X., Yang Q., Ding T., Xu J., Yan Z., Men Y., Xin W, & Xu H. (2019). Retracted Article: Gm5820, an antisense RNA of FGF1, suppresses FGF1 expression at the posttranscriptional level to inactivate the ERK/STAT3 pathway and alleviates neuropathic pain in mice. RSC Advances, 9(49), 28364-28376.

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Immunohistochemical Analysis of M. hyorhinis and TNF-α in PCa Tissues

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PCa tissue microarrays (TMA, BC19021a, US Biomax, Rockville, MD) were deparaffined and the antigens were retrieved using a pressure cooker. The tissues were washed three times in PBS, permeabilized with PBS containing 0.1 % Triton X-100 for 10 min, and then incubated with 2 % BSA for 1 h. The tissues were incubated overnight at 4 °C with mouse anti-M. hyorhinis (P70 surface antigen) (1:500, EMZ104, Kerafast, Boston, MA) and rabbit anti-TNF-α (1:400, 8184, Cell Signaling). After washing three times in PBS, cells were incubated with Alexa Fluor 594 or 488 coupled secondary antibodies for 1 h, washed three times in PBS, and then mounted using Vectashield antifade mounting medium with DAPI (H-1200, Vector Laboratories, Burlingame, CA). Images were taken by whole slide scanning from UCLA Translational Pathology Core Laboratory and using Aperio ImageScope software (Leica Biosystems).

Kim J.K., Chang I., Jung Y., Kaplan Z., Hill E.E., Taichman R.S, & Krebsbach P.H. (2023). Mycoplasma hyorhinis infection promotes TNF-α signaling and SMAC mimetic-mediated apoptosis in human prostate cancer. Heliyon, 9(10), e20655.

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Immunofluorescent Labeling of Brain Tissue

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Similar procedures to process the tissue were followed as described for standard IHC. In addition, 50μm sections were washed in TBS (pH 7.2) for 10 min prior to antigen retrieval with sodium citrate buffer (pH 6.0) incubated for 30 min at 80°C. Tissues were permeabilized using 0.2% Triton X in TBS for 15 min at RT, blocked using 4% normal goat serum (NGS) in 0.1% Tween/TBS for 1h at RT, prior to exposure to: i) chicken anti-GFAP (Novus Biologicals USA, Littleton, CO; 1:1500) and rabbit anti-PV (ABcam PLC; 1:1000) or rabbit anti-TNF-α (Cell Signaling Technology, Inc; 1:250) in 4% NGS in TBS overnight at 4°C; or ii) chicken anti-α-tubulin (Abcam PLC; 1:1000) and rabbit anti-PSD95 (Proteintech Group, Inc. Rosemont, IL; 1:200). Following incubation with primary antibodies and washes, tissues were incubated in the dark for 2h at RT exposed to goat anti-chicken IgY Alexa Fluor 488 and goat anti-rabbit Alexa Fluor 568 emitting a green and red fluorescence signal, respectively (Thermo Fisher Scientific, Inc, Waltham, MA). Following exposure to secondary antibodies, tissues were incubated for 5 min in 4’,6-daamidino-2-phenylindole (DAPI; 1μg/mL) in TBS, washed in TBS, and mounted and dried for 30 min prior to coverslip with ProLong Gold Antifade Mountant (Molecular Probes, Life Technologies Corp., Carlsbad, CA).

Chavez-Valdez R., Emerson P., Martin L.J., Goffigan-Holmes J., Kirkwood A, & Northington F.J. (2018). Delayed injury of hippocampal interneurons after neonatal hypoxia-ischemia and therapeutic hypothermia in a murine model.. Hippocampus, 28(8), 617-630.

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Rosemary Extract Neuroprotective Mechanisms

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Rosemary extracts (Kunming Pharmaceutical Co., China.) are the crucial active constituents extracted from Rosmarinus officinalis Linn, containing 60% carnosic acid. RE were dissolved in 1% Tween-80 to prepare a suspension with the concentration of 100 mg/mL. Rabbit anti-BDNF, rabbit anti-Iba1 and rabbit anti-IL-1β were purchased from Abcam (Shanghai, China). Rabbit anti-TNF-α, rabbit anti-p-NF κ B p65, rabbit anti-NF κ B p65, rabbit anti-AKT, rabbit anti-p-AKT 473, β-actin and horseradish peroxidase conjugated anti-rabbit IgG were acquired from Cell Signaling Technology (Boston, MA, United States). Enzyme-linked immunosorbent assay (ELISA) kits were obtained from RD, Bio-Techne (Emeryville, CA, United States). E.Z.N.A. stool DNA Kit was purchased from Omega Bio-tek (Norcross, GA, United States), AxyPrep DNA Gel Extraction Kit was acquired from Axygen Biosciences (Union City, CA, United States).

Guo Y., Xie J., Li X., Yuan Y., Zhang L., Hu W., Luo H., Yu H, & Zhang R. (2018). Antidepressant Effects of Rosemary Extracts Associate With Anti-inflammatory Effect and Rebalance of Gut Microbiota. Frontiers in Pharmacology, 9, 1126.

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Neuroprotective Effects of Chondroitin Sulfate

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CS from shark cartilage was purchased from Jiaxing Hengjie Biopharmaceutical Co., Ltd. (Jiaxing, China). CS (Mw = 4.2 kDa) used in this paper was prepared in our laboratory [15 (link)]. LiCl was purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Cation-exchange resin and ultrafiltration membrane with MWCO of 1000 Da and 5000 Da were from Sangon Biotech Co., Ltd. (Shanghai, China). The amyloid-β (1-42) peptide was purchased from GL Biochem Ltd. (Shanghai, China). AchE, ChAT, and Na⁺/K⁺-ATPase assay kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). GSH-Px and MDA assay kits were obtained from Beyotime Institute of Biotechnology (Jiangsu, China). Rabbit anti-TNF-α, anti-IL-6, anti-IL-1β, anti-p-p38MAPK, anti-p-Erk1/2, anti-p-NF-κB, anti-p-GSK3β (Ser9), and anti-PP2A antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA). Rabbit anti-p-tau (Ser396/Ser404), anti-tau, and anti-GSK3β antibodies were purchased from Abcam (Cambridge, MA, USA). Rabbit anti-IκBa, anti-p-IκBa, and anti-Lamin B were purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (Beijing, China). The TUNEL assay kit was purchased from Roche (Basel, Switzerland).

Gao D., Li P., Gao F., Feng Y., Li X., Li D., Li Y, & Xiao Y. (2022). Preparation and Multitarget Anti-AD Activity Study of Chondroitin Sulfate Lithium in AD Mice Induced by Combination of D-Gal/AlCl3. Oxidative Medicine and Cellular Longevity, 2022, 9466166.

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Microglial Cell Immunostaining Protocol

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Microglial cells after 48 h exposure to TNF-α with or without inhibitors were fixed in 4% PFA-methanol free solution (ThermoFisher, Milan, Italy), and permeabilized with 0.05% Triton X-100 (Bio-Rad Laboratories, Milan, Italy) for 3–5 min at room temperature. The cells were then rinsed with DPBS, and incubated with the primary antibodies overnight at 4 °C. The primary antibodies used were mouse ant-Iba1 (1:1000, #66,827–1-Ig, Proteintech, DBA, Milan, Italy), rabbit anti-TMEM119 (1:1000, #66,948–1-Ig Proteintech, DBA, Milan, Italy), mouse anti-iNOS (1:1000, #18,985–1-AP Proteintech, DBA, Milan, Italy), and rabbit anti-TNF-α (1:1000, #3707; Cell Signaling Technology, DBA, Milan, Italy). Secondary antibodies conjugated to Alexa 488 (1:250, #A-11029, Invitrogen, Milan, Italy) or 594 (1:250, #R37117, Invitrogen, Milan, Italy) fluorochromes were used to detect the primary antibodies. DAPI (Fluoroshield Mounting Medium with DAPI, #Ab104139 Abcam, Cambridge, UK) was used to visualize the nuclei and count cells.

Serafini S., Ferretti G., Monterosso P., Angiolillo A., Di Costanzo A, & Matrone C. (2024). TNF-α Levels Are Increased in Patients with Subjective Cognitive Impairment and Are Negatively Correlated with β Amyloid-42. Antioxidants, 13(2), 216.

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Western Blotting Analysis of Cellular and Tissue Proteins

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Western blotting analysis was performed as previously described.^{25 (link)} Cellular and mouse tissue proteins were exposed to radioimmunoprecipitation assay (RIPA) buffer (50 mmol/L Tris‐HCl [pH 7.6], 150 mmol/L NaCl, 1% NP‐40, 0.5% sodium deoxycholate, and 0.1% SDS), homogenized on ice and centrifuged. The supernatants were resolved via 10% SDS‐PAGE and were transferred to PVDF membranes (Millipore). The membranes were blocked with Tris‐buffered saline (TBS) containing 5% non‐fat milk and were incubated with the following primary antibodies: rabbit anti‐IRF7 (sc‐9083; 1:200; Santa Cruz), mouse anti‐PCNA (#2586; 1:1000; Cell Signaling Technology), rabbit anti‐Cyclin D1 (#2978; 1:1000; Cell Signaling Technology), rabbit anti‐NF‐κB p65 (#4764; 1:1000; Cell Signaling Technology), rabbit anti‐phosphorylated‐NF‐κB p65 (ser536) (BS4138; 1:1000; Bioworld), mouse anti‐IκBα (#4814; 1:1000; Cell Signaling Technology), mouse anti‐phosphorylated IκBα (BS4105; 1:1000; Bioword), rabbit anti‐TNFα (#3707; 1:1000; Cell Signaling Technology), goat anti‐IL‐6 (AF‐406‐NA; 1:500; R&D System), goat anti‐Collagen type I (sc‐8784; 1:1000; Santa Cruz), goat anti‐Collagen type III (sc‐8781; 1:1000; Santa Cruz), rabbit anti‐ATF3 (sc‐188; 1:200; Santa Cruz), and mouse anti‐GAPDH (MB001; 1:10 000; Bioworld).

Huang L., Zhang S., Zhang P., Zhang X., Zhu L., Chen K., Gao L., Zhang Y., Kong X., Tian S., Zhang X, & Li H. (2014). Interferon Regulatory Factor 7 Protects Against Vascular Smooth Muscle Cell Proliferation and Neointima Formation. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(5), e001309.

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