重组
RabMAb

Anti-SLP76 (phospho Y145)抗体[EP2853Y] (ab75829)

概述

  • 产品名称
    Anti-SLP76 (phospho Y145)抗体[EP2853Y]
    参阅全部 SLP76 一抗
  • 描述
    兔单克隆抗体[EP2853Y] to SLP76 (phospho Y145)
  • 特异性
    This antibody detects SLP76 phosphorylated on tyrosine 145.
  • 经测试应用
    适用于: ICC/IF, WB, Flow Cytmore details
    不适用于: IHC-P
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide within Human SLP76 aa 100-200 (phospho Y145). The exact sequence is proprietary.
    Database link: Q13094

  • 阳性对照
    • Jurkat cell lysates treated with pervanadate.
  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • 纯度
    Tissue culture supernatant
  • 克隆
    单克隆
  • 克隆编号
    EP2853Y
  • 同种型
    IgG
  • 研究领域

相关产品

应用

Our Abpromise guarantee covers the use of ab75829 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
ICC/IF 1/500.
WB 1/1000 - 1/10000. Predicted molecular weight: 60 kDa.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • 应用说明
    Is unsuitable for IHC-P.
  • 靶标

    • 功能
      Involved in T-cell antigen receptor mediated signaling.
    • 组织特异性
      Highly expressed in spleen, thymus and peripheral blood leukocytes. Highly expressed also in T-cell and monocytic cell lines, expressed at lower level in B-cell lines. Not detected in fibroblast or neuroblastoma cell lines.
    • 序列相似性
      Contains 1 SAM (sterile alpha motif) domain.
      Contains 1 SH2 domain.
    • 结构域
      The SH2 domain mediates interaction with SHB.
    • 翻译后修饰
      Phosphorylated after T-cell receptor activation by ZAP-70.
    • 细胞定位
      Cytoplasm.
    • Information by UniProt
    • 数据库链接
    • 别名
      • 76 kDa tyrosine phosphoprotein antibody
      • CG8697 antibody
      • LCP 2 antibody
      • LCP2 antibody
      • LCP2_HUMAN antibody
      • Lymphocyte cytosolic protein 2 antibody
      • SH2 domain containing leukocyte protein 76 KD antibody
      • SH2 domain containing leukocyte protein of 76kD antibody
      • SH2 domain containing leukocyte protein of 76kDa antibody
      • SH2 domain-containing leukocyte protein of 76 kDa antibody
      • SLP 76 antibody
      • SLP 76 tyrosine phosphoprotein antibody
      • SLP-76 antibody
      • SLP-76 tyrosine phosphoprotein antibody
      • SLP76 antibody
      • SLP76 tyrosine phosphoprotein antibody
      see all

    Anti-SLP76 (phospho Y145) antibody [EP2853Y] 图像

    • ab75829 staining SLP76 in Jurkat by Immunocytochemistry. Tissue was fixed with 4% paraformaldehyde. Samples were incubated with primary antibody (4.5 μg/ml). ab150077 AlexaFluor®488 Goat anti-Rabbit (1/1000) was used as the secondary antibody. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 2.5 μg/ml and DAPI were used as counter stains.

      Confocal image showing the expression was increased after treatment with pervanadate (1mM, 30min) and then decreased after treatment with the Lambda Protein Phosphatase 31℃ for 2h

    • All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/5000 dilution

      Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
      Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates. treated with pervanadate at 50nM for 5 min.
      Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates. Treated with pervanadate at 50nM for 5 min. Then the membrane was incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 60 kDa
      Observed band size : 76 kDa (why is the actual band size different from the predicted?)

      Blocking buffer 5% NFDM/TBST. Diluting buffer 5% NFDM/TBST

    • ab75829 staining SLP76 (phospho Y145) antibody in both treated (50mM of pervanadate for 5 minutes) and untreated Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000.

      DAPI was used as a nuclear counterstain.

    • All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/200000 dilution

      Lane 1 : Jurkat cell lysates un-treated
      Lane 2 : Jurkat cell lysates treated with pervanadate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 60 kDa
      Observed band size : 76 kDa (why is the actual band size different from the predicted?)
    • Dot blot analysis of SLP76 (phospho Y145) phospho peptide (Lane 1), SLP76 Non-phospho peptide (Lane 2), labelling SLP76 (phospho Y145) with ab75829 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.

      Blocking and diluting buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Flow cytometric analysis of permeabilized Jurkat cells, untreated (green) or pervanadate treated (red) using ab75829 at a 1/100 dilution. Also, pervanadate Jurkat cells using the same antibody preincubated with phospho SLP76 peptide (blue) or non phospho SLP76 peptide (orange).

    Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)参考文献

    This product has been referenced in:
    • Yang CY  et al. Dual-specificity phosphatase 14 (DUSP14/MKP6) negatively regulates TCR signaling by inhibiting TAB1 activation. J Immunol 192:1547-57 (2014). Read more (PubMed: 24403530) »
    • Gill T & Levine AD Mitochondria-derived hydrogen peroxide selectively enhances T cell receptor-initiated signal transduction. J Biol Chem 288:26246-55 (2013). Read more (PubMed: 23880762) »

    See all 2 Publications for this product

    Product Wall

    Application
    Western blot
    Loading amount
    100000 cells
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - whole cell (jurkats)
    Specification
    jurkats
    Treatment
    TCR activation with OKT3 and anti CD8
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Dr. Tejpal Gill

    Verified customer

    提交于 May 13 2014

    Thank you for contacting us.

    Your credit note ID is 23976.

    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used i...

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    Thank you for your enquiry regarding ab75829 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

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    Thank you for your inquiry.

    The dilution factors for each application listed on the product datasheet for ab75829 are what we currently recommended, it is still relevant.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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