RabMAb

Anti-Src (phospho Y529)抗体[Y232] (ab32078)

概述

  • 产品名称Anti-Src (phospho Y529)抗体[Y232]
    参阅全部 Src 一抗
  • 描述
    兔单克隆抗体[Y232] to Src (phospho Y529)
  • 特异性ab32078 will detect Src phosphorylation on Tyrosine 529 of both isoforms. The antibody immunogen shares 93% homology with Fyn and Yes and 85% homology with Fgr. Therefore, it is likely that the antibody will cross-react with these proteins. However, this is just based on BLAST results and no experiments were performed.
  • 经测试应用适用于: IP, WB, Dot blot, ICC/IFmore details
    不适用于: Flow Cyt or IHC
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Src.
    (Peptide available as ab179556)

  • 阳性对照
    • WB: HeLa and A431 cell lysates ICC/IF: HeLa and A431 cells
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

性能

相关产品

应用

Our Abpromise guarantee covers the use of ab32078 in the following tested applications.

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

应用 Ab评论 说明
IP 1/50.
WB 1/5000 - 1/10000. Predicted molecular weight: 60 kDa.Can be blocked with Src (phospho Y529) peptide (ab179556).
Dot blot 1/2000.
ICC/IF 1/50 - 1/500.
  • 应用说明Is unsuitable for Flow Cyt or IHC.
  • 靶标

    • 功能Non-receptor protein tyrosine kinase that plays pivotal roles in numerous cellular processes such as proliferation, migration, and transformation. In concert with PTK2B, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. Once it is recruited to the activated integrins, by PTK2B, it phosphorylates CBL which in turn induces the activation and recruitment of phosphatidylinositol 3-kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. Promotes energy production in osteoclasts by activating mitochondrial cytochrome C oxidase. Phosphorylates RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284' and CBL on 'Tyr-731'. Enhances DDX58/RIG-I-elicited antiviral signaling.
    • 序列相似性Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
      Contains 1 protein kinase domain.
      Contains 1 SH2 domain.
      Contains 1 SH3 domain.
    • 翻译后修饰Dephosphorylated at Tyr-530 by PTPRJ (By similarity). Phosphorylated on Tyr-530 by c-Src kinase (CSK). The phosphorylated form is termed pp60c-src. Dephosphorylated by PTPRJ at Tyr-419. Normally maintained in an inactive conformation with the SH2 domain engaged with Tyr-530, the SH3 domain engaged with the SH2-kinase linker, and Tyr-419 dephosphorylated. Dephosphorylation of Tyr-530 as a result of protein tyrosine phosphatase (PTP) action disrupts the intramolecular interaction between the SH2 domain and Tyr-530, Tyr-419 can then become autophosphorylated, resulting in SRC activation. Phosphorylation of Tyr-530 by CSK allows this interaction to reform, resulting in SRC inactivation.
      S-nitrosylation is important for activation of its kinase activity.
    • 细胞定位Cell membrane. Mitochondrion inner membrane.
    • Information by UniProt
    • 数据库链接
    • 形式This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
    • 别名
      • ASV antibody
      • Avian sarcoma virus antibody
      • c SRC antibody
      • CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA antibody
      • cSrc antibody
      • EC 2.7.10.2 antibody
      • Neuronal CSRC tyrosine specific protein kinase antibody
      • Neuronal SRC antibody
      • Oncogene SRC antibody
      • OTTHUMP00000174476 antibody
      • OTTHUMP00000174477 antibody
      • p60 Src antibody
      • p60-Src antibody
      • p60Src antibody
      • pp60c src antibody
      • pp60c-src antibody
      • pp60csrc antibody
      • Proto oncogene tyrosine protein kinase Src antibody
      • Proto-oncogene c-Src antibody
      • Proto-oncogene tyrosine-protein kinase Src antibody
      • Protooncogene SRC antibody
      • Protooncogene SRC Rous sarcoma antibody
      • Src antibody
      • SRC Oncogene antibody
      • SRC proto oncogene non receptor tyrosine kinase antibody
      • SRC_HUMAN antibody
      • SRC1 antibody
      • Tyrosine kinase pp60c src antibody
      • Tyrosine protein kinase SRC 1 antibody
      • Tyrosine protein kinase SRC1 antibody
      • v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog antibody
      • V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian) antibody
      • v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian antibody
      see all

    Anti-Src (phospho Y529) antibody [Y232] 图像



    • Predicted band size : 60 kDa

      WB analysis. Lane 1:Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates 15ug and Lane 2:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H2O2 at 10mM for 1 h. whole cell lysates 15ug and Lane 3:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H2O2 at 10mM for 1 h. whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Primary ab used at 1:30,000 dilution. 5% NFDM/TBST  was used as Blocking buffer and Diluting buffer. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as a Secondary ab at 1:20,000 dilution. The Exposure time was 30 seconds.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling Src (phospho Y529) with ab32078 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.

      The green staining was increased in the H2O2 (10mM, 1 hour) treated HeLa cells when compared with HeLa cells without treatment. After LP treatment, the green signaling was obviously decreased.

      For the pan antibody, there was no great difference after H2O2 (10 mM, 1 hour) or EGF (100 ng/mL, 10 minutes) + LP treatment.

      The data showed mostly Cytoplasm and Membran staining for Ab32078.

    • Dot Blot analysis of Lane 1: Src (pY529) phospho peptide and Lane 2: Src non-phospho peptide labeling Src (phospho Y529) with ab32078 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.

    • All lanes : Anti-Src (phospho Y529) antibody [Y232] (ab32078) at 1/10000 dilution

      Lane 1 : Untreated A431 cells
      Lane 2 : EGF treated A431 cells
      Lane 3 : EGF and alkaline phosphatase treated A431 cells


      Predicted band size : 60 kDa
      Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    • Dot Blot analysis on immunogen phospho peptide (A) and non phospho peptide (B) using 1/2000 ab32078.

    Anti-Src (phospho Y529) antibody [Y232] (ab32078)参考文献

    This product has been referenced in:
    • Li M  et al. Arenobufagin, a bufadienolide compound from toad venom, inhibits VEGF-mediated angiogenesis through suppression of VEGFR-2 signaling pathway. Biochem Pharmacol 83:1251-60 (2012). Read more (PubMed: 22305746) »
    • Su L  et al. P-glycoprotein regulates blood-testis barrier dynamics via its effects on the occludin/zonula occludens 1 (ZO-1) protein complex mediated by focal adhesion kinase (FAK). Proc Natl Acad Sci U S A : (2011). WB . Read more (PubMed: 22106313) »

    See all 3 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 100000 cells
    Gel Running Conditions Reduced Denaturing
    Sample Human Cell lysate - whole cell (Jurkat)
    Specification Jurkat
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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    Abcam user community

    Verified customer

    提交于 Feb 20 2014

    Thank you for contacting us.

    We have not tested these antibodies in Drosophila, but BLAST searches showed low homology for both antibodies, 40-50%, so I would not predict reactivity.

    I hope this information is helpful to you. Please d...

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    Thank you for contacting us.

    For ab4817, ab32078 the immuinogen peptide are unfortunately not available.

    I have inquired about ab47411 and will let you know when I hear back form the lab.

    Thank you for your patience.In t...

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