Anti-SHC antibody [EP332Y] (ab33770)
- Datasheet
- Specific References (2)
- Protocols
Overview
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Product nameAnti-SHC antibody [EP332Y]
See all SHC primary antibodies -
DescriptionRabbit monoclonal [EP332Y] to SHC
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Host speciesRabbit
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SpecificityThis antibody detects p46, p52 and p66 -three isoforms of SHC.
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Tested applicationsSuitable for: Flow Cyt, ICC/IF, IHC-P, WBmore details
Unsuitable for: IP -
Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide within Human SHC aa 500-600. The exact sequence is proprietary.
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Positive control
- MCF 7 cell lysate
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General notes
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.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEP332Y
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Positive Controls
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 |
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| Flow Cyt | 1/140. For unpurified use at 1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| ICC/IF | 1/200. | |
| IHC-P | 1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| WB | 1/1000 - 1/2000. Predicted molecular weight: 63 kDa. For unpurified use at 1/2000. |
Target
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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.
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Tissue specificityWidely expressed. Expressed in neural stem cells but absent in mature neurons.
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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.
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Post-translational
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
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Database links
- Entrez Gene: 6464 Human
- Entrez Gene: 20416 Mouse
- Entrez Gene: 85385 Rat
- Omim: 600560 Human
- SwissProt: P29353 Human
- SwissProt: P98083 Mouse
- SwissProt: Q5M824 Rat
- Unigene: 433795 Human
see all -
Alternative names
- FLJ26504 antibody
- p66 antibody
- p66SHC antibody
see all
Images
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![Flow Cytometry - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770-240636-33770-Flow.jpg)
Flow Cytometry - Anti-SHC antibody [EP332Y] (ab33770)Overlay histogram showing MCF-7 cells stained with ab33770(red line) at 1/140 dilution. The cells were fixed with 2% paraformaldehyde. The secondary antibody used was a FITC conjugated goat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (green line) was rabbit monoclonal IgG used under the same conditions.
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![Immunocytochemistry/ Immunofluorescence - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770-240634-33770-IF.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-SHC antibody [EP332Y] (ab33770)ab33770 staining SHC in the MCF-7 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/200). ab150077(1/500) an Alexa Fluor®488-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770-240631-33770-IHC2.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770-240630-33770-IHC1.jpg)
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.
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![Western blot - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770-240629-33770-WB2.jpg)
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 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Western blot - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770-240628-33770-WB1.jpg)
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 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Western blot - Anti-SHC antibody [EP332Y] (ab33770)](https://www.abcam.com/ps/products/33/ab33770/Images/ab33770.jpg)
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)
References
This product has been referenced in:
- Krochmal M et al. Identification of novel molecular signatures of IgA nephropathy through an integrative -omics analysis. Sci Rep 7:9091 (2017). Read more (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 . Read more (PubMed: 19797472) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"