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Tnf r1 h 5

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TNF-R1 (H-5) is an antibody product developed by Santa Cruz Biotechnology for research purposes. It targets the tumor necrosis factor receptor 1 (TNFR1) protein. The antibody can be used to detect and study TNFR1 expression and function in various experimental systems, but its specific applications and intended uses are not provided in this factual description.

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Lab products found in correlation

Tnf r1 c25c1, by Cell Signaling Technology (2 mentions) Traf2 f 2, by Santa Cruz Biotechnology (2 mentions) Tradd 37 tradd, by BD (2 mentions) Bond max automated immunohistochemistry vision biosystem, by Leica (1 mentions) Epitope retrieval solution 2 ph9, by Leica (1 mentions) Bond primary antibody diluent, by Leica (1 mentions) Bond polymer refine detection kit, by Leica (1 mentions) Peroxidase conjugated goat anti mouse igg h l antibody, by Jackson ImmunoResearch (1 mentions) β actin, by Merck Group (1 mentions) Ripk1, by BD (1 mentions) Phospho iκbα ser32 36, by Cell Signaling Technology (1 mentions) Ripk1 38 rip, by BD (1 mentions) Lamin a c e1, by Santa Cruz Biotechnology (1 mentions) Flag tag (flag), by Fujifilm (1 mentions) Peroxidase conjugated anti mouse igg h l antibody, by Jackson ImmunoResearch (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Ecl select reagent, by GE Healthcare (1 mentions) P iκbα ser 32 36, by Santa Cruz Biotechnology (1 mentions) Alliance ld2, by Uvitec (1 mentions) Gapdh 6c5, by Santa Cruz Biotechnology (1 mentions)

5 protocols using tnf r1 h 5

1

Immunohistochemistry Protocol for ETV7/TNFR1

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On a few clinical samples from Santa Chiara Hospital, APSS, Trento, IHC was performed in a Bond Max Automated Immunohistochemistry Vision Biosystem (Leica Microsystems, Wetzlar, Germany) using the Bond Polymer Refine Detection kit (DS9800; Leica Biosystems, Wetzlar, Germany) as previously described [67 (link)]. Briefly, 3-µm-thick sections were prepared from formalin-fixed paraffin-embedded tissue blocks, deparaffinized, pre-treated with epitope retrieval solution 2 (pH9; Leica Biosystems) at 100 °C for 20 min, and then incubated for 30 min with primary antibodies (ETV7/TEL2, E-1, 1:50; TNFR1, H-5, 1:50, Santa Cruz Biotechnology) diluted in Bond Primary Antibody Diluent (AR9352; Leica Biosystems). All the slides were reviewed by a pathologist. Informed consent for use of tissue materials for research purposes was approved by the local ethics committee (APSS, Trento, Italy) and was obtained from all subjects.
Meškytė E.M., Pezzè L., Bartolomei L., Forcato M., Bocci I.A., Bertalot G., Barbareschi M., Oliveira-Ferrer L., Bisio A., Bicciato S., Baltriukienė D, & Ciribilli Y. (2023). ETV7 reduces inflammatory responses in breast cancer cells by repressing the TNFR1/NF-κB axis. Cell Death & Disease, 14(4), 263.
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2

Antibody Identification for Western Blotting

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Antibodies reactive to β-actin (clone: AC-15, catalog number: A5441; Sigma-Aldrich), the DYKDDDDK tag (FLAG) (clone: 1E6, code number: 012-22384; Fujifilm Wako Pure Chemical Corporation), RIPK1 (clone: 38/RIP, material number: 610458; BD Biosciences, San Jose, CA, USA), TNF-R1 (H-5, catalog number: sc-8436; Santa Cruz Biotechnology, Dallas, TX, USA), TNF-R1 (C25C1, catalog number: #3736; Cell Signaling Technology, Danvers, MA, USA), TRADD (clone: 37/TRADD, material number: 610572; BD Biosciences), and TRAF2 (F-2, catalog number: sc-136999; Santa Cruz Biotechnology) were used as primary antibodies for Western blotting. A peroxidase-conjugated goat anti-mouse IgG (H + L) antibody (code number: 115-035-146) and peroxidase-conjugated goat anti-rabbit IgG (H + L) antibody (code number: 111-035-144) were obtained from Jackson ImmunoResearch Laboratories (West Groove, PA, USA), which were used as secondary antibodies for Western blotting.
Vu Q.V., Sayama S., Ando M, & Kataoka T. (2024). Sesquiterpene Lactones Containing an α-Methylene-γ-Lactone Moiety Selectively Down-Regulate the Expression of Tumor Necrosis Factor Receptor 1 by Promoting Its Ectodomain Shedding in Human Lung Adenocarcinoma A549 Cells. Molecules, 29(8), 1866.
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3

Immunoblotting of Apoptosis-related Proteins

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Primary antibodies for γ1-actin (2F3; FUJIFILM Wako Pure Chemical Corporation), FLAG (1E6; FUJIFILM Wako Pure Chemical Corporation), ICAM-1 (15.2; Leinco Technologies, Inc., St. Louis, MO, USA), RelA (F-6; Santa Cruz Biotechnology, Dallas, TX, USA), lamin A/C (E-1; Santa Cruz Biotechnology), GAPDH (6C5; Santa Cruz Biotechnology), IκBα (25/IkBa/MAD-3; BD Biosciences, San Jose, CA, USA), phospho-IκBα (Ser32/36) (5A5; Cell Signaling Technology, Danvers, MA, USA), TNF-R1 (H-5; Santa Cruz Biotechnology), TNF-R1 (C25C1; Cell Signaling Technology), RIPK1 (38/RIP; BD Biosciences), TRADD (37/TRADD; BD Biosciences), and TRAF2 (F-2; Santa Cruz Biotechnology) were used. A peroxidase-conjugated anti-mouse IgG(H + L) antibody (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) and peroxidase-conjugated anti-rabbit IgG(H + L) antibody (Jackson ImmunoResearch Laboratories) were used as secondary antibodies.
Kusagawa E., Okuda C., Yamaguchi R., Nakano K., Miyake Y, & Kataoka T. (2022). Cucurbitacin B Down-Regulates TNF Receptor 1 Expression and Inhibits the TNF-α-Dependent Nuclear Factor κB Signaling Pathway in Human Lung Adenocarcinoma A549 Cells. International Journal of Molecular Sciences, 23(13), 7130.
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4

Western Blot Analysis of Cell Signaling

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Total protein cell extracts were obtained by lysing the cells with RIPA buffer supplemented with 1x protease inhibitors (PI) (Roche, Milan, Italy). Proteins were quantified using the BCA method (Pierce, ThermoFisher Scientific), and then 30–50 µg of proteins were loaded on 8–12% polyacrylamide gels for SDS-PAGE. After the separation, the proteins were transferred on a nitrocellulose membrane (Amersham, Merck,) which was probed over-night at 4 °C with specific antibodies diluted in 1–3% skimmed milk-PBS-0.1% Tween solution: TNFR1 (H-5, Santa Cruz Biotechnologies, DBA, Milan, Italy), STAT3 (124H6, Cell Signaling Technologies, Euroclone, Milan, Italy), pSTAT3 (Y705, Cell Signaling Technologies), HSP70 (C92F3A-5, Santa Cruz Biotechnologies), TEL2 (E-1, Santa Cruz Biotechnologies), GAPDH (6C5, Santa Cruz Biotechnologies), H3 (Abcam), pIκBα (Ser32/36, Santa Cruz Biotechnologies). Detection was performed with ECL Select Reagent (GE Healthcare, Cytiva) using UVITec Alliance LD2 (UVITec Cambridge, UK) imaging system.
Meškytė E.M., Pezzè L., Bartolomei L., Forcato M., Bocci I.A., Bertalot G., Barbareschi M., Oliveira-Ferrer L., Bisio A., Bicciato S., Baltriukienė D, & Ciribilli Y. (2023). ETV7 reduces inflammatory responses in breast cancer cells by repressing the TNFR1/NF-κB axis. Cell Death & Disease, 14(4), 263.
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5

Western Blot Analysis of Signaling Proteins

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The preparation of cell lysates and western blotting were performed as described previously. 16, 17 Protein samples (30 μg) in sample buffer (62.5 mM Tris, 2% SDS, 10% glycerol, 0.003% bromophenol blue and 288 mM 2-mercaptoethanol) were boiled at 100 °C for 5 min, fractionated by SDS-polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes. The membranes were incubated with primary antibodies specific to γ1-actin (2F3; Wako Pure Chemical Industries), FLAG (1E6; Wako Pure Chemical Industries), IκBα (clone 25; BD Biosciences, San Jose, CA, USA), IκB kinase β (D30C6; Cell Signaling Technology, Danvers, MA, USA), RelA (C-20; Santa Cruz Biotechnology, Santa Cruz, CA, USA), RIP1 (38/RIP; BD Biosciences), TNF-R1 (H-5; Santa Cruz Biotechnology), TRADD (37/ TRADD; BD Biosciences), TRAF2 (C-20; Santa Cruz Biotechnology) and horseradish peroxidase-linked secondary antibodies (Jackson ImmunoResearch Labora-tories, West Grove, PA, USA). Blots were analyzed by ImageQuant LAS 4000 mini (GE Healthcare).
Quach H.T., Tanigaki R., Yokoigawa J., Yamada Y., Niwa M., Hirano S., Shiono Y., Kimura K.I, & Kataoka T. (2017). Allantopyrone A interferes with multiple components of the TNF receptor 1 complex and blocks RIP1 modifications in the TNF-α-induced signaling pathway. The Journal of antibiotics, 70(9).
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