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使用敲除细胞株进行验证重组RabMAb

重组Anti-PBK/SPK抗体[EP2520Y] (ab75987)

Submit a review Submit a question References (9)
Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
  • Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
  • Anti-PBK/SPK antibody [EP2520Y] (ab75987)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2520Y] to PBK/SPK
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-PBK/SPK抗体[EP2520Y]
    参阅全部 PBK/SPK 一抗

  • 描述

    兔单克隆抗体[EP2520Y] to PBK/SPK

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IP, IHC-Pmore details
    不适用于: Flow Cyt or ICC/IF

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide within Human PBK/SPK (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: HEK-293T, HeLa and HAP1 cell lysates. IHC-P: Human tonsil tissue

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab75987于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/100000 - 1/500000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IP
1/30.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.
说明
WB
1/100000 - 1/500000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IP
1/30.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.
应用说明
Is unsuitable for Flow Cyt or ICC/IF.

靶标

  • 功能

    Phosphorylates MAP kinase p38. Seems to be active only in mitosis. May also play a role in the activation of lymphoid cells. When phosphorylated, forms a complex with TP53, leading to TP53 destabilization and attenuation of G2/M checkpoint during doxorubicin-induced DNA damage.

  • 组织特异性

    Expressed in the testis and placenta. In the testis, restrictedly expressed in outer cell layer of seminiferous tubules.

  • 序列相似性

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. MAP kinase kinase subfamily.
    Contains 1 protein kinase domain.

  • 翻译后修饰

    Phosphorylated; in a cell-cycle dependent manner at mitosis.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Cancer/testis antigen 84 antibody
      • CT84 antibody
      • Epididymis luminal protein 164 antibody
      • FLJ14385 antibody
      • HEL164 antibody
      • Lymphokine activated killer T cell originated protein kinase antibody
      • Lymphokine-activated killer T-cell-originated protein kinase antibody
      • MAPKK like protein kinase antibody
      • MAPKK-like protein kinase antibody
      • Nori 3 antibody
      • Nori-3 antibody
      • Nori3 antibody
      • PBK antibody
      • PDZ binding kinase antibody
      • PDZ-binding kinase antibody
      • Serine/threonine protein kinase antibody
      • Spermatogenesis related protein kinase antibody
      • Spermatogenesis-related protein kinase antibody
      • SPK antibody
      • T LAK cell originated protein kinase antibody
      • T-LAK cell-originated protein kinase antibody
      • TOPK antibody
      • TOPK_HUMAN antibody
      see all

    图片

    • Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
      Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PBK knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Human tonsil tissue lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - loading control, ab7291, observed at 52 kDa.

      ab75987 was shown to recognize PBK when PBK knockout samples were used, along with additional cross-reactive bands. Wild-type and PBK knockout samples were subjected to SDS-PAGE. ab75987 diluted to 1/100,000 and ab7291 (loading control to alpha Tubulin) diluted to 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] (ab75987)

      Immunohistochemical analysis of paraffin-embedded human tonsil using ab75987 at 1/100 dilution. Antigen retrieval was heat mediated with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
      Western blot - Anti-PBK/SPK antibody [EP2520Y] (ab75987)
      All lanes : Anti-PBK/SPK antibody [EP2520Y] (ab75987) at 1/100000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : PBK knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 36 kDa
      Observed band size: 40 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab75987 was shown to react with PBK/SPK in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266827 (knockout cell lysate ab257575) was used. Wild-type HEK-293T and PBK knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab75987 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 100000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Anti-PBK/SPK antibody [EP2520Y] (ab75987)
      Anti-PBK/SPK antibody [EP2520Y] (ab75987)

    文献 (9)

    发表研究结果有使用 ab75987？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab75987 被引用在 9 文献中.

    • Li F  et al. Identification of potential biomarkers in cancer testis antigens for glioblastoma. Am J Transl Res 15:799-816 (2023). PubMed: 36915736
    • Ma H  et al. PBK promotes aggressive phenotypes of cervical cancer through ERK/c-Myc signaling pathway. J Cell Physiol 236:2767-2781 (2021). PubMed: 33184870
    • Mao P  et al. PDZ-Binding Kinase-Dependent Transcriptional Regulation of CCNB2 Promotes Tumorigenesis and Radio-Resistance in Glioblastoma. Transl Oncol 13:287-294 (2020). PubMed: 31874375
    • Park JH  et al. TOPK inhibition accelerates oxidative stress-induced granulosa cell apoptosis via the p53/SIRT1 axis. Int J Mol Med 46:1923-1937 (2020). PubMed: 32901866
    • Ma H  et al. PBK, targeted by EVI1, promotes metastasis and confers cisplatin resistance through inducing autophagy in high-grade serous ovarian carcinoma. Cell Death Dis 10:166 (2019). PubMed: 30778048
    • Huo X  et al. Clinical and Expression Significance of AKT1 by Co-expression Network Analysis in Endometrial Cancer. Front Oncol 9:1147 (2019). PubMed: 31781484
    • Dou X  et al. PBK/TOPK mediates geranylgeranylation signaling for breast cancer cell proliferation. Cancer Cell Int 15:27 (2015). Human . PubMed: 25745361
    • Lei B  et al. PBK/TOPK expression correlates with mutant p53 and affects patients' prognosis and cell proliferation and viability in lung adenocarcinoma. Hum Pathol 46:217-24 (2015). PubMed: 25466965
    • Guo Z  et al. Proteomics strategy to identify substrates of LNX, a PDZ domain-containing E3 ubiquitin ligase. J Proteome Res 11:4847-62 (2012). PubMed: 22889411

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