使用敲除细胞株进行验证

Anti-TPMT抗体[AT2E7] (ab122984)

概述

  • 产品名称
    Anti-TPMT抗体[AT2E7]
    参阅全部 TPMT 一抗
  • 描述
    小鼠单克隆抗体[AT2E7] to TPMT
  • 经测试应用
    适用于: Flow Cyt, ICC/IF, WB, ELISAmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Recombinant full length protein, corresponding to amino acids 1-245 of Human TPMT purified from E. coli (NP_000358).

  • 阳性对照
    • Hep3B cell lysate.

性能

应用

Our Abpromise guarantee covers the use of ab122984 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
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 28 kDa.
ELISA Use at an assay dependent concentration.

靶标

  • 功能
    Catalyzes the S-methylation of thiopurine drugs such as 6-mercaptopurine.
  • 疾病相关
    Defects in TPMT are the cause of thiopurine S-methyltransferase deficiency (TPMT deficiency) [MIM:610460]. TPMT is an enzyme involved in the normal metabolic inactivation of thiopurine drugs. These drugs are generally used as immunosupressants or cytotoxic drugs and are prescribed for a variety of clinical conditions including leukemia, autoimmune disease and organ transplantation. Patients with intermediate or no TPMT activity are at risk of toxicity after receiving standard doses of thiopurine drugs and it is shown that inter-individual differences in response to these drugs are largely determined by genetic variation at the TPMT locus.
  • 序列相似性
    Belongs to the methyltransferase superfamily. TPMT family.
  • 细胞定位
    Cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • HGNC:12014 antibody
    • S adenosyl L methionine thiopurine S methyltransferase antibody
    • Thiopurine methyltransferase antibody
    • Thiopurine S methyltransferase antibody
    • Thiopurine S-methyltransferase antibody
    • TPMT antibody
    • TPMT_HUMAN antibody
    see all

图片



  • Predicted band size : 28 kDa

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: TPMT knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HepG2 whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab122984 observed at 28 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab122984 was shown to recognize TPMT in wild-type HAP1 cells as signal was lost at the expected MW in TPMT knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TPMT knockout samples were subjected to SDS-PAGE. Ab122984 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Anti-TPMT antibody [AT2E7] (ab122984) at 1/1000 dilution + Hep3B lysate at 40 µg

    Secondary
    goat anti-mouse secondary antibody conjugated to HRP
    Developed using the ECL technique

    Predicted band size : 28 kDa


  • Predicted band size : 28 kDa

    The Recombinant Protein (50ng) and Cell lysates (40ug) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human TPMT antibody (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

    Lane 1 : Recombinant Protein
    Lane 2 : HepG2 cell lysate

  • Immunocytochemistry/Immunofluorescence analysis of U87MG cells labelling TPMT with ab122984 at 1/100. An Alexa Fluor® 488-conjugated goat anti-mouse IgG was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

  • Flow Cytometry analysis of U87MG cells labelling TPMT with ab122984, staining at 2-5 µg/1x106 cells (red). A 488-conjugated goat anti-mouse IgG was used as the secondary antibody. Black - Isotype control, mouse IgG.

文献

ab122984 has not yet been referenced specifically in any publications.

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