Anti phospho shc tyr239 240

Manufactured by Cell Signaling Technology

Anti-phospho-SHC (Tyr239/240) is a laboratory reagent used for the detection and analysis of phosphorylated SHC proteins. It is a specific antibody that recognizes the Tyr239/240 phosphorylation sites on SHC proteins. This product can be used in various analytical techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the activation and signaling pathways involving SHC proteins.

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Lab products found in correlation

Anti cldn1, by Thermo Fisher Scientific (1 mentions) Anti phospho p44 42 erk thr202 tyr204, by Cell Signaling Technology (1 mentions) Anti ocln, by Thermo Fisher Scientific (1 mentions) Cl xposure film, by Thermo Fisher Scientific (1 mentions) Sds page gel, by Bio-Rad (1 mentions) West femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Anti erk, by Thermo Fisher Scientific (1 mentions) Supersignal west pico plus chemiluminescent, by Thermo Fisher Scientific (1 mentions) Anti egfr, by Cell Signaling Technology (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Complete mini protease inhibitor, by Roche (1 mentions) Anti β actin, by R&D Systems (1 mentions) Anti shc, by Cell Signaling Technology (1 mentions) Odyssey clx imaging system, by LI COR (1 mentions) Fluorescence conjugated secondary antibody, by LI COR (1 mentions) Lysis buffer, by Merck Group (1 mentions) Protease and phosphatase inhibitor, by Merck Group (1 mentions)

Spelling variants of this product

Anti-Shc (3 citations) Anti-Phospho-Shc (2 citations) Phospho-SHC (2 citations)

2 protocols using anti phospho shc tyr239 240

Western Blotting Protocol for Protein Analysis

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All cells were lysed in 1% NP40 buffer (150 mM NaCl, 50 mM Tris-HCl pH 8, 10% glycerol, 2 mM EDTA) supplemented with 1 mM cOmplete Mini protease inhibitors (Roche) and 1 mM sodium orthovanadate (ThermoFisher). Proteins were separated on a 4–20% SDS-PAGE gel (BioRad) and transferred to polyvinylidene difluoride (PVDF) membrane. Membrane was incubated in 5% BSA and 0.1% Tween-20 in PBS for 1 hr, incubated overnight at 4°C with primary antibodies (1:10000 anti-β-actin, Santa Cruz #sc-47778; 1:1000 anti-SHC, BD Transduction #610878; 1:1000 anti-ERK, Invitrogen #13–6200; 1:1000 anti-phospho-SHC (Tyr239/240), Cell Signaling #2434; 1:2000 anti-phospho-p44/42 ERK (Thr202/Tyr204), Cell Signaling #4370; 1:1000 anti-EGFR, Cell Signaling #2232; 1:700 anti-phospho-Thr/Tyr, Cell Signaling #9381; 1:500 anti-CLDN1, Invitrogen #37–4900; 1:350 anti-OCLN, Invitrogen #33–1500), then incubated for 1 hr at room temperature with horseradish peroxidase-conjugated secondary antibodies (1:10000 anti-rabbit, Cell Signaling #7074; 1:10000 anti-mouse, Cell Signaling #7076). Membrane was added SuperSignal West Pico PLUS Chemiluminescent or West Femto Maximum Sensitivity substrate (ThermoFisher) and exposed to CL-XPosure film (ThermoFisher).

So C.W., Sourisseau M., Sarwar S., Evans M.J, & Randall G. (2023). Roles of epidermal growth factor receptor, claudin-1 and occludin in multi-step entry of hepatitis C virus into polarized hepatoma spheroids. PLOS Pathogens, 19(12), e1011887.

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Western Blot Analysis of Phospho-Shc

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Liver tissues were homogenized in the lysis buffer (MilliporeSigma) containing protease and phosphatase inhibitors (MilliporeSigma). Thirty micrograms of proteins were separated proteins subjected to 4%–20% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membranes. After blocking, the membranes were probed with primary antibodies as indicated at 4°C overnight, then with fluorescence conjugated secondary antibodies (LI-COR) for 1 h at room temperature. Images were visualized and analyzed using Odyssey CLx Imaging System (LI-COR).
To detect phospho-Shc and Shc in PBMC, automated Western blot analysis was conducted using the Jess™ capillary western system (ProteinSimple) following the instructions. Total 0.75 μg of protein was separated and immobilized in electrophoretic capillaries provided by the manufacturer. Primary antibodies (1:50 to 1:100) were applied for 1 h. After probing with appropriate secondary antibodies for another hour, the signal was acquired and quantified with Compass software (ProteinSimple).
Anti-phospho-Shc (Tyr239/240), and anti-Shc were from Cell Signaling Inc.; anti-β-Actin was from R&D Systems.

Jiang J.X., Tomilov A., Montgomery C., Hui C.K., Török N.J, & Cortopassi G. (2021). Shc inhibitor idebenone ameliorates liver injury and fibrosis in dietary NASH in mice. Journal of biochemical and molecular toxicology, 35(10), e22876.

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