Validated using a knockout cell lineRabMAb

Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)

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Key features and details

  • Rabbit monoclonal [EP1465Y] to Serine/threonine-protein kinase 4/MST-1
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y]
    See all Serine/threonine-protein kinase 4/MST-1 primary antibodies

  • Description

    Rabbit monoclonal [EP1465Y] to Serine/threonine-protein kinase 4/MST-1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Serine/threonine-protein kinase 4/MST-1 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Jurkat, Ramos and HeLa cell lysate and mouse spleen, rat spleen and human urinary bladder tissues. ICC/IF: Raw264.7 cells. IHC-P:Human gastric carcinoma. IP: Jurkat cell lysate. Flow Cyt: HeLa cells.

  • General notes

    Previously labelled as Serine/threonine-protein kinase 4

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

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab51134 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
Flow Cyt 1/50.
WB 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 56 kDa).
IHC-P 1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

We strongly recommend that customers perform an antigen retrieval step.
ICC/IF 1/100 - 1/250.
IP 1/30 - 1/100.

Target

  • Function

    Stress-activated, pro-apoptotic kinase which, following caspase-cleavage, enters the nucleus and induces chromatin condensation followed by internucleosomal DNA fragmentation. Key component of the Hippo signaling pathway which plays a pivotal role in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein MST1/MST2, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. Phosphorylation of YAP1 by LATS2 inhibits its translocation into the nucleus to regulate cellular genes important for cell proliferation, cell death, and cell migration. MST1/MST2 are required to repress proliferation of mature hepatocytes, to prevent activation of facultative adult liver stem cells (oval cells), and to inhibit tumor formation (By similarity). Phosphorylates 'Ser-14' of histone H2B (H2BS14ph) during apoptosis. Phosphorylates FOXO3 upon oxidative stress, which results in its nuclear translocation and cell death initiation.

  • Tissue specificity

    Ubiquitously expressed.

  • Sequence similarities

    Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.
    Contains 1 protein kinase domain.
    Contains 1 SARAH domain.

  • Post-translational
    modifications

    Autophosphorylated on serine and threonine residues.

  • Cellular localization

    Cytoplasm. Nucleus. The caspase-cleaved form cycles between the nucleus and cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • Kinase responsive to stress antibody
      • Krs2 antibody
      • Mammalian STE20 like protein kinase 1 antibody
      • Mammalian STE20-like protein kinase 1 antibody
      • Mammalian sterile 20 like 1 antibody
      • MST-1 antibody
      • MST1 antibody
      • Serine/threonine kinase 4 antibody
      • Serine/threonine protein kinase Krs 2 antibody
      • Serine/threonine-protein kinase 4 antibody
      • Serine/threonine-protein kinase Krs-2 antibody
      • STE20 like kinase MST1 antibody
      • STE20-like kinase MST1 antibody
      • STK4 antibody
      • STK4_HUMAN antibody
      • TIIAC antibody
      • YSK3 antibody
      see all

    Images

    • Western blot - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Western blot - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      All lanes : Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : MST-1 knockout HeLa cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : Ramos cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 56 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab51134 observed at 52 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab51134 Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] was shown to specifically react with MST-1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265442 (knockout cell lysate ab258215) was used. Wild-type and MST-1 knockout samples were subjected to SDS-PAGE. ab51134 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.

       

    • Western blot - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Western blot - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      All lanes : Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134) at 1/50000 dilution

      Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
      Lane 2 : Mouse spleen
      Lane 3 : Rat spleen

      Lysates/proteins at 1/20 dilution per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 56 kDa



      Diluting and blocking buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Immunocytochemistry/ Immunofluorescence - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)

      ab51134 staining Serine/threonine-protein kinase 4/MST-1 in Raw264.7 (mouse abelson murine leukemia virus-induced tumor) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a concentration of 1/1000. ab7291 anti-Tubulin (mouse mAb) (1/1000) and ab150120 AlexaFluor®594 Goat anti-Mouse secondary (1/1000)  were used as counterstains for primary antibody ab51134 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

      Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
      Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)

      ab51134 staining Serine/threonine-protein kinase 4/MST-1 in human gastric carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/50. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

      Negative control 1: PBS in place of primary antibody.

    • Flow Cytometry - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Flow Cytometry - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)

      Flow Cytometry analysis of HeLa cells labelling Serine/theronine-protein kinase 4/MST-1 with purified ab51134 at a dilution of 1/50 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • Immunoprecipitation - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Immunoprecipitation - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)

      ab51134 immunoprecipitating Serine/threonine-protein kinase 4/MST-1. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/30 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.

      Lane 1: Jurkat (human acute T cell leukemia) whole cell lysate (10ug)
      Lane 2: Jurkat (human acute T cell leukemia) whole cell lysate
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab51134 in Jurkat (human acute T cell leukemia) whole cell lysate

    • Western blot - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Western blot - Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134)
      Anti-Serine/threonine-protein kinase 4/MST-1 antibody [EP1465Y] (ab51134) at 1/10000 dilution + HeLa (human cervix adenocarcinoma) whole cell lysate at 20 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 56 kDa



      Diluting and blocking buffer: 5% NFDM/TBST

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (18)

    Publishing research using ab51134? Please let us know so that we can cite the reference in this datasheet.

    ab51134 has been referenced in 18 publications.

    • Shang Y  et al. Over-expressed MST1 impaired spatial memory via disturbing neural oscillation patterns in mice. Genes Brain Behav N/A:e12678 (2020). PubMed: 32468668
    • Hu L  et al. Tri-ortho-cresyl phosphate (TOCP) induced ovarian failure in mice is related to the Hippo signaling pathway disruption. Reprod Toxicol 83:21-27 (2019). PubMed: 30439503
    • Cheng Z  et al. Cardiac-specific Mst1 deficiency inhibits ROS-mediated JNK signalling to alleviate Ang II-induced cardiomyocyte apoptosis. J Cell Mol Med 23:543-555 (2019). PubMed: 30338935
    • Yoshida M  et al. miR-7977 inhibits the Hippo-YAP signaling pathway in bone marrow mesenchymal stromal cells. PLoS One 14:e0213220 (2019). PubMed: 30835743
    • Du X  et al. Hippo/Mst signalling couples metabolic state and immune function of CD8a+ dendritic cells. Nature 558:141-145 (2018). IP . PubMed: 29849151
    View all Publications for this product

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