Overview

  • Product name

    Anti-PBK/SPK antibody [EPR21983]
    See all PBK/SPK primary antibodies
  • Description

    Rabbit monoclonal [EPR21983] to PBK/SPK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PBK/SPK aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q96KB5

  • Positive control

    • IHC-P: Human, rat and mouse testis tissue. Human gastric cancer tissue. WB: HeLa, HepG2, SW480, MCF7, A431, U-87 MG, Hepa1-6, C6, PC-12 and RAW 264.7 whole cell lysate. Human and rat testis lysate. Wild-type HAP1 cell lysate. IP: PBK/SPK IP in HepG2 whole cell lysate. Flow Cyt: HeLa and HepG2 cells.
  • General notes

     This product was previously labelled as PBK

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab236872 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
WB 1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/500.
IP 1/30.

Target

  • Function

    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.
  • Tissue specificity

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

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. MAP kinase kinase subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

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

  • Alternative names

    • 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

Images

  • All lanes : Anti-PBK/SPK antibody [EPR21983] (ab236872) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : PBK/SPK knockout HAP1 cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 36 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID:23547718; PMID:25909225).

    ab236872 was shown to specifically react with PBK/SPK in wild-type HAP1 cells as signal was lost in PBK/SPK knockout cells. Wild-type and PBK/SPK knockout samples were subjected to SDS-PAGE. ab236872 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of human testis (PMID:16982762; PMID:25909225) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-PBK/SPK antibody [EPR21983] (ab236872) at 1/1000 dilution

    Lane 1 : SW480 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
    Lane 3 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate
    Lane 4 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
    Lane 5 : Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate
    Lane 6 : Human testis lysate
    Lane 7 : Rat testis lysate
    Lane 8 : C6 (rat glial tumor glial cell), whole cell lysate
    Lane 9 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
    Lane 10 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 36 kDa
    Observed band size: 38 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lanes 1-4 & 8-9: 15 seconds; Lanes 5 & 10: 3 minutes; Lane 6: 70 seconds; Lane 7: 3 seconds.

    The molecular weight observed is consistent with what has been described in the literature (PMID:23547718; PMID:25909225).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling PBK/SPK with ab236872 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • PBK/SPK was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab236872 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236872 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
    Lane 1: HepG2 whole cell lysate 10 µg (Input).
    Lane 2: ab236872 IP in HepG2 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236872 in HepG2 whole cell lysate.
    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 10 seconds.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of rat testis (PMID:16982762; PMID:25909225) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of mouse testis (PMID:16982762; PMID:25909225) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling PBK/SPK with ab236872 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human gastric cancer (PMID:26894977; PMID:27898655) is observed. Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cell line labeling PBK/SPK with ab236872 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

References

ab236872 has not yet been referenced specifically in any publications.

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