Overview

  • Product name

    Anti-SPAK antibody [EPR6394]
    See all SPAK primary antibodies
  • Description

    Rabbit monoclonal [EPR6394] to SPAK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human SPAK aa 400-500.
    Database link: Q9UEW8

  • Positive control

    • SH-SY5Y, 293T, Jurkat, BxPC3 and HepG2 whole cell lysate (ab7900); Human colon tissue
  • General notes

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

     

     This product was previously labelled as STK39

     

    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 ab128894 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 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 59 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/10 - 1/100.

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

  • Application notes
    Is unsuitable for ICC or IP.
  • Target

    • Function

      May act as a mediator of stress-activated signals.
    • Tissue specificity

      Predominantly expressed in brain and pancreas followed by heart, lung, kidney, skeletal muscle, liver, placenta and testis.
    • Sequence similarities

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

      PAPA box (proline-alanine repeats) may target the kinase to a specific subcellular location by facilitating interaction with intracellular proteins such as actin or actin-like proteins.
    • Cellular localization

      Cytoplasm. Nucleus. Nucleus when caspase-cleaved.
    • Information by UniProt
    • Database links

    • Alternative names

      • DCHT antibody
      • DKFZp686K05124 antibody
      • OTTHUMP00000165175 antibody
      • PASK antibody
      • Proline alanine rich STE20 related kinase antibody
      • Serine threonine kinase 39 (STE20/SPS1 homolog yeast) antibody
      • Serine threonine kinase 39 antibody
      • Serine/threonine protein kinase 39 antibody
      • Serine/threonine-protein kinase 39 antibody
      • Small intestine SPAK like kinase antibody
      • SPAK antibody
      • Ste 20 related kinase antibody
      • Ste-20-related kinase antibody
      • Ste20 like protein kinase antibody
      • STE20/SPS1 homolog antibody
      • STE20/SPS1 related proline alanine rich protein kinase antibody
      • STE20/SPS1-related proline-alanine-rich protein kinase antibody
      • STK 39 antibody
      • Stk39 antibody
      • STK39_HUMAN antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: SPAK knockout HAP1 whole cell lysate (20 µg)
      Lane 3: Jurkat whole cell lysate (20 µg)
      Lane 4: MCF7 whole cell lysate (20 µg)


      Lanes 1 - 4: Merged signal (red and green). Green - ab128894 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab128894 was shown to specifically react with SPAK in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when SPAK knockout samples were uexamined. Wild-type and SPAK knockout samples were subjected to SDS-PAGE. Ab128894 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-SPAK antibody [EPR6394] (ab128894) at 1/1000 dilution

      Lane 1 : SH-SY5Y cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : BxPC3 cell lysate
      Lane 5 : HepG2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 59 kDa
      Observed band size: 65 kDa why is the actual band size different from the predicted?

    • ab128894, at 1/50 dilution, staining SPAK in paraffin embedded Human colon tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Wolley MJ  et al. In Primary Aldosteronism, Mineralocorticoids Influence Exosomal Sodium-Chloride Cotransporter Abundance. J Am Soc Nephrol 28:56-63 (2017). Human . Read more (PubMed: 27381844) »
    See 1 Publication for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A549)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    20 µg
    Specification
    A549
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Dec 16 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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