Recombinant
RabMAb

Recombinant Anti-SHC antibody [EP332Y] (ab33770)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP332Y] to SHC
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-SHC antibody [EP332Y]
    See all SHC primary antibodies

  • Description

    Rabbit monoclonal [EP332Y] to SHC

  • Host species

    Rabbit

  • Specificity

    This antibody detects p46, p52 and p66 -three isoforms of SHC.

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human SHC aa 500-600. The exact sequence is proprietary.

  • Positive control

    • MCF7 whole cell lysate (ab3871) can be used as a positive control in WB. MCF 7 cell lysate

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab33770 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
WB 1/1000 - 1/2000. Predicted molecular weight: 63 kDa.

For unpurified use at 1/2000.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.

    • Target

    • Function

      Signaling adapter that couples activated growth factor receptors to signaling pathways. Participates in a signaling cascade initiated by activated KIT and KITLG/SCF. Isoform p46Shc and isoform p52Shc, once phosphorylated, couple activated receptor tyrosine kinases to Ras via the recruitment of the GRB2/SOS complex and are implicated in the cytoplasmic propagation of mitogenic signals. Isoform p46Shc and isoform p52Shc may thus function as initiators of the Ras signaling cascade in various non-neuronal systems. Isoform p66Shc does not mediate Ras activation, but is involved in signal transduction pathways that regulate the cellular response to oxidative stress and life span. Isoform p66Shc acts as a downstream target of the tumor suppressor p53 and is indispensable for the ability of stress-activated p53 to induce elevation of intracellular oxidants, cytochrome c release and apoptosis. The expression of isoform p66Shc has been correlated with life span (By similarity). Participates in signaling downstream of the angiopoietin receptor TEK/TIE2, and plays a role in the regulation of endothelial cell migration and sprouting angiogenesis.

    • Tissue specificity

      Widely expressed. Expressed in neural stem cells but absent in mature neurons.

    • Sequence similarities

      Contains 1 PID domain.
      Contains 1 SH2 domain.

    • Domain

      In response to a variety of growth factors, isoform p46Shc and isoform p52Shc bind to phosphorylated Trk receptors through their phosphotyrosine binding (PID) and/or SH2 domains. The PID and SH2 domains bind to specific phosphorylated tyrosine residues in the Asn-Pro-Xaa-Tyr(P) motif of the Trk receptors. Isoform p46Shc and isoform p52Shc are in turn phosphorylated on three tyrosine residues within the extended proline-rich domain. These phosphotyrosines act as docking site for GRB2 and thereby are involved in Ras activation.

    • Post-translational
      modifications

      Phosphorylated by activated epidermal growth factor receptor. Phosphorylated in response to FLT4 and KIT signaling. Isoform p46Shc and isoform p52Shc are phosphorylated on tyrosine residues of the Pro-rich domain. Isoform p66Shc is phosphorylated on Ser-36 by PRKCB upon treatment with insulin, hydrogen peroxide or irradiation with ultraviolet light (By similarity). Tyrosine phosphorylated in response to FLT3 signaling (By similarity). Tyrosine phosphorylated by activated PTK2B/PYK2 (By similarity). Tyrosine phosphorylated by ligand-activated ALK. Tyrosine phosphorylated by ligand-activated PDGFRB. Tyrosine phosphorylated by TEK/TIE2. May be tyrosine phosphorylated by activated PTK2/FAK1; tyrosine phosphorylation was seen in an astrocytoma biopsy, where PTK2/FAK1 kinase activity is high, but not in normal brain tissue. Isoform p52Shc dephosphorylation by PTPN2 may regulate interaction with GRB2.

    • Cellular localization

      Cytoplasm; Mitochondrion matrix. Localized to the mitochondria matrix. Targeting of isoform p46Shc to mitochondria is mediated by its first 32 amino acids, which behave as a bona fide mitochondrial targeting sequence. Isoform p52Shc and isoform p66Shc, that contain the same sequence but more internally located, display a different subcellular localization and Mitochondrion. In case of oxidative conditions, phosphorylation at 'Ser-36' of isoform p66Shc, leads to mitochondrial accumulation.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • FLJ26504 antibody
      • p66 antibody
      • p66SHC antibody
      • SH2 domain protein C1 antibody
      • SHC (Src homology 2 domain containing) transforming protein 1 antibody
      • SHC 1 antibody
      • SHC A antibody
      • SHC adaptor protein 1 antibody
      • Shc antibody
      • SHC transforming protein 1 antibody
      • SHC transforming protein antibody
      • SHC-transforming protein 1 antibody
      • SHC-transforming protein 3 antibody
      • SHC-transforming protein A antibody
      • SHC1 antibody
      • SHC1_HUMAN antibody
      • SHCA antibody
      • Src homology 2 domain-containing-transforming protein C1 antibody
      see all

    Images

    • Western blot - Anti-SHC antibody [EP332Y] (ab33770)
      Western blot - Anti-SHC antibody [EP332Y] (ab33770)
      All lanes : Anti-SHC antibody [EP332Y] (ab33770) at 1/1000 dilution

      Lane 1 : C6 Cell lysate
      Lane 2 : RAW264.7 Cell lysate
      Lane 3 : NIH/3T3 Cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

      Predicted band size: 63 kDa



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHC antibody [EP332Y] (ab33770)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHC antibody [EP332Y] (ab33770)

      ab33770 staining SHC in Human renal carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/300). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHC antibody [EP332Y] (ab33770)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHC antibody [EP332Y] (ab33770)

      ab33770 staining SHC in Human stomach tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/300). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

    • Western blot - Anti-SHC antibody [EP332Y] (ab33770)
      Western blot - Anti-SHC antibody [EP332Y] (ab33770)
      All lanes : Anti-SHC antibody [EP332Y] (ab33770) at 1/1000 dilution

      Lane 1 : MCF-7 cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

      Predicted band size: 63 kDa



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-SHC antibody [EP332Y] (ab33770)
      Western blot - Anti-SHC antibody [EP332Y] (ab33770)
      Anti-SHC antibody [EP332Y] (ab33770) at 1/2000 dilution (unpurified) + MCF 7 cell lysate

      Predicted band size: 63 kDa
      Observed band size: 66 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 46 kDa (possible isoform), 52 kDa (possible isoform)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (5)

    Publishing research using ab33770? Please let us know so that we can cite the reference in this datasheet.

    ab33770 has been referenced in 5 publications.

    • Benkafadar N  et al. ROS-Induced Activation of DNA Damage Responses Drives Senescence-Like State in Postmitotic Cochlear Cells: Implication for Hearing Preservation. Mol Neurobiol N/A:N/A (2019). PubMed: 30693443
    • Zeng M  et al. Chinese yam extract and adenosine attenuated LPS-induced cardiac dysfunction by inhibiting RAS and apoptosis via the ER-mediated activation of SHC/Ras/Raf1 pathway. Phytomedicine 61:152857 (2019). PubMed: 31035040
    • Jiang W  et al. Klotho inhibits PKCa/p66SHC-mediated podocyte injury in diabetic nephropathy. Mol Cell Endocrinol 494:110490 (2019). PubMed: 31207271
    • Krochmal M  et al. Identification of novel molecular signatures of IgA nephropathy through an integrative -omics analysis. Sci Rep 7:9091 (2017). PubMed: 28831120
    • Yang HC  et al. The PPARgamma agonist pioglitazone ameliorates aging-related progressive renal injury. J Am Soc Nephrol 20:2380-8 (2009). WB ; Rat . PubMed: 19797472

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