Ab131205

Manufactured by Abcam

Sourced in United States

Ab131205 is a synthetic peptide corresponding to a region within human HMGB1 protein. The product is intended for use in ELISA and other immunoassay applications.

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Lab products found in correlation

3 protocols using ab131205

Caerulein-Induced Mitochondrial Dynamics Analysis

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Caerulein (CAE) was purchased from Sigma-Aldrich (St. Louis, MO, United States). Rabbit monoclonal antibodies against OPA1 (ab157457, Abcam Plc, Cambridge, United Kingdom), DRP1 (D6C7 rabbit mAb; Cell Signaling), LC3B [LC3B antibody (2775)]; Cell Signaling Technology, MA, United States), p62 [SQSTM1 (P-15): sc-10117; Santa Cruz Biotechnology, Santa Cruz, CA, United States)], VMP1 (D1y3E, Cell Signaling Technology, MA, United States); mouse monoclonal antibodies against Parkin (P6248, Sigma-Aldrich, St. Louis, MO, United States), V5 [13202 V5-Tag (D3H8Q) rabbit mAB-Cell Signaling)], β-actin [β-actin (C4): sc-47778; Santa Cruz Biotechnology, Santa Cruz, CA, United States)], beta tubulin (ab131205, Abcam Plc, Cambridge, United Kingdom); rabbit anti-goat antibody (Santa Cruz Biotechnology, Santa Cruz, CA, United States); goat policlonal antibodies against VDAC-1 (D-16 sc-32063; Santa Cruz Biotechnology, Santa Cruz, CA, United States); rabbit anti-mouse antibody [(315-035-048) Jackson InmunoResearch, Baltimore Pike, United States]; goat anti-rabbit antibody [(GAR):170-5046; Bio-Rad, CA, United States]. Other reagents, enzymes, and chemicals were of reagent grade and also from Sigma-Aldrich.

Vanasco V., Ropolo A., Grasso D., Ojeda D.S., García M.N., Vico T.A., Orquera T., Quarleri J., Alvarez S, & Vaccaro M.I. (2021). Mitochondrial Dynamics and VMP1-Related Selective Mitophagy in Experimental Acute Pancreatitis. Frontiers in Cell and Developmental Biology, 9, 640094.

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Quantitative Immunoblotting of Cellular Proteins

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About 10⁷ cells were lysed in 100 μl of RIPA buffer with a protease inhibitor cocktail and EDTA (Thermo Fisher Scientific). Protein concentration was determined using Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). Proteins were separated on 4%–15% Mini‐PROTEAN TGX Stain‐free Gel (Bio‐Rad). After electrophoresis, proteins were transferred onto a 0.45‐μm PVDF Low Fluorescence membrane (Bio‐Rad). Membranes were blocked using 5% non‐fat milk and incubated with 1:2000 anti‐CD45 antibody (610266, BD Biosciences), 1:2000 anti‐SOCS2 antibody (#2779, Cell Signaling Technology), or 1:10000 anti‐β‐tubulin antibody (ab131205, Abcam). After washing, membranes were incubated with horseradish peroxidase (HRP) conjugated with 1:10,000 anti‐mouse secondary antibody (A9917, Sigma Aldrich) or 1:40,000 anti‐rabbit secondary antibody (ab97051, Abcam). Immunoreactive protein bands were detected with Clarity Western ECL Substrate for HRP (Bio‐Rad) on Chemidoc Imaging System (Bio‐Rad). The abundance of target protein was assessed in reference to β‐tubulin loading control using Image Lab 6.0.1 software (Bio‐Rad). Each experiment was conducted in three biological replicates.

Drobna‐Śledzińska M., Maćkowska‐Maślak N., Jaksik R., Kosmalska M., Szarzyńska B., Lejman M., Sędek Ł., Szczepański T., Taghon T., Van Vlierberghe P., Witt M, & Dawidowska M. (2022). Multiomics to investigate the mechanisms contributing to repression of PTPRC and SOCS2 in pediatric T‐ALL: Focus on miR‐363‐3p and promoter methylation. Genes, Chromosomes & Cancer, 61(12), 720-733.

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Immunoblotting Analysis of Signaling Pathways

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Primary antibodies for TLR-4 (#30002), NOX4 (#21860) and β-actin (#47778) and secondary antibodies rabbit anti-goat IgG-HRP (#2768), mouse anti-rabbit IgG-HRP (#2357), and goat anti-mouse IgG-HRP (#2005) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary and secondary antibodies for IκB kinase α (IKKα) (#11930), p-IKKα/β (Ser176/180) (#2697), inhibitor of κBα (IκBα) (#4814), p-IκBα (Ser32) (#2859), NF-κB p65 (#8242) and p-NF-κB p65 (Ser536) (#3033) were obtained from Cell Signaling Technology (Boston, MA, USA). Primary antibodies for NOX1 (ab46545) and NOXO1 (ab131205) were purchased from Abcam Inc. (Cambridge, MA, USA). (-)-Epicatechin was obtained from Sigma Chemical Co. (St Louis, MO, USA), and fructose ( purity ≥99.5%) was obtained from Droguería Saporiti (Buenos Aires, Argentina). Commercial rat chow was purchased from Gepsa-Feeds (Buenos Aires, Argentina).

Prince PD ., Rodríguez Lanzi C ., Fraga CG , & Galleano M . (2019). Dietary (-)-epicatechin affects NF-κB activation and NADPH oxidases in the kidney cortex of high-fructose-fed rats. Food & function, 10(1).

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