Anti shc

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-Shc is a primary antibody that recognizes the Shc (Src homology 2 domain containing) adaptor protein. Shc proteins play a role in signal transduction pathways by mediating the activation of downstream signaling cascades.

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Lab products found in correlation

Close spelling variants of this product

Rabbit anti-Shc (3 citations) Anti-phospho-SHC (Tyr239/240), (2 citations) Anti-Shc antibody (2 citations) Anti-Phospho-Shc (2 citations)

3 protocols using anti shc

Western Blotting Analysis of Neuroblastoma Cells

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Neuroblastoma cell lines were seeded in a 10-cm dish at a final density of 1.5×10⁶ cells and allowed to attach overnight. The cells were then treated with DMSO or experimental compounds. After 48–72 h, cells were lysed in CHAPS lysis buffer with protease and phosphatase inhibitors (Roche, Basel, Switzerland), and 25 or 50 μg of total lysate were boiled for 5–10 min in CHAPS and sample buffer. Western blotting was performed using the following primary antibodies: anti-ALK (Cell Signaling Technology, Danvers, MA, USA), anti-Phospho-ALK (Cell Signaling Technology), anti-Shc (Cell Signaling Technology), anti-Phospho-Shc (Cell Signaling Technology), anti-PARP (Cell Signaling Technology), anti-cleaved Caspase-3 (Cell Signaling Technology), anti-GAPDH (Fuji Film-Wako Chemicals), and anti-Actin (Merck). Secondary antibodies used were as follows: HRP-linked anti-rabbit IgG (Cell Signaling Technology) and HRP-linked anti-mouse IgG (Cell Signaling Technology). The proteins were visualized using ImageQuant LAS 4000mini with enhanced chemiluminescence reagents (Thermo Fisher Scientific).

Ota Y., Yoda H., Inoue T., Watanabe T., Shinozaki Y., Takatori A, & Nagase H. (2021). Targeting anaplastic lymphoma kinase (ALK) gene alterations in neuroblastoma by using alkylating pyrrole-imidazole polyamides. PLoS ONE, 16(9), e0257718.

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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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Antibody Validation for Signaling Pathways

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The following antibodies were used: anti–phospho-ERK, anti-fibroblast growth factor receptor substrate 2 (FRS2), anti-p85 PI3-kinase, and anti–penta-His tag from Santa Cruz Biotechnology (Dallas, TX, USA); anti–phospho-FGFR (Tyr653/654), anti-FGFR1, anti-KDR, anti–phospho-FRS2, anti–phospho-Src family (Tyr416), anti-Src, anti-growth factor receptor-bound protein 2 (Grb2), anti-Shc, anti–phospho-Akt (Ser473), and anti-Akt from Cell Signaling Technology (Danvers, MA, USA); anti–phospho-tyrosine (clone 4G10) from MilliporeSigma (Burlington, MA, USA); anti-ERK2 from Bioss Antibodies (Woburn, MA, USA); anti-FLAG M2 antibody from MilliporeSigma; anti-HA.11 epitope tag antibody from BioLegend (San Diego, CA, USA); anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from AbClon (Seoul, Republic of Korea); horseradish peroxidase–conjugated goat anti-mouse IgG and rabbit IgG from KOMA Biotech (Seoul, Republic of Korea). The anti-PTK7 antibody was previously described by Shin et al. (20 (link)).

Shin W.S., Lee H.W, & Lee S.T. (2019). Catalytically inactive receptor tyrosine kinase PTK7 activates FGFR1 independent of FGF. The FASEB Journal, 33(11), 12960-12971.

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