Product nameAnti-SHC (phospho S36) antibody [6E10]
See all SHC primary antibodies
DescriptionMouse monoclonal [6E10] to SHC (phospho S36)
SpecificityThis antibody recognizes the 66kDa form of SHC protein phosphorylated on Ser36. It does not cross react with the non-phosphorylated form of SHC or with unrelated phosphorylation sites.
Tested applicationsSuitable for: WB, ELISA, IHC-P, ICC/IF, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic phosphopeptide corresponding to amino acids surrounding the Ser36 phosphorylation site of human SHC.
- HeLa cells stimulated with EGF
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.1% Sodium azide
Constituents: Sucrose, Polyethylene glycol, PBS
Concentration information loading...
Purification notesPurified from ascites.
Our Abpromise guarantee covers the use of ab54518 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 63 kDa.|
|ELISA||Use a concentration of 0.05 µg/ml.|
|IHC-P||Use at an assay dependent concentration. PubMed: 24273694|
|ICC/IF||Use at an assay dependent concentration. PubMed: 21799744|
|Flow Cyt||Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionSignaling 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 specificityWidely expressed. Expressed in neural stem cells but absent in mature neurons.
Sequence similaritiesContains 1 PID domain.
Contains 1 SH2 domain.
DomainIn 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.
modificationsPhosphorylated 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 localizationCytoplasm; 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
- FLJ26504 antibody
- p66 antibody
- p66SHC antibody
All lanes : Anti-SHC (phospho S36) antibody [6E10] (ab54518) at 1 µg/ml
Lane 1 : HeLa cells stimulated with EGF for 0 mins.
Lane 2 : HeLa cells stimulated with EGF for 1 min.
Lane 3 : HeLa cells stimulated with EGF for 5 mins.
Lane 4 : HeLa cells stimulated with EGF for 15 mins.
Lane 5 : HeLa cells stimulated with EGF for 30 mins.
Lane 6 : HeLa cells stimulated with EGF for 60 mins.
Lysates/proteins at 15 µl per lane.
Developed using the ECL technique.
Predicted band size: 63 kDa
Observed band size: 67 kDa why is the actual band size different from the predicted?
Overlay histogram showing MCF7 cells stained with ab54518 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54518, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This product has been referenced in:
- Zhang T et al. p66Shc is associated with hydrogen peroxide-induced oxidative stress in preimplantation sheep embryos. Mol Reprod Dev 86:342-350 (2019). Read more (PubMed: 30636355) »
- Halon-Golabek M et al. hmSOD1 gene mutation-induced disturbance in iron metabolism is mediated by impairment of Akt signalling pathway. J Cachexia Sarcopenia Muscle N/A:N/A (2018). WB . Read more (PubMed: 29380557) »