Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PYK2 antibody [YE353] - BSA and Azide free (ab228477)

Overview

  • Product name

    Anti-PYK2 antibody [YE353] - BSA and Azide free
    See all PYK2 primary antibodies
  • Description

    Rabbit monoclonal [YE353] to PYK2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human PYK2 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Ramos, Jurkat and RAW264.7 cell lysates and mouse and rat brain tissue lysates. IHC-P: Human, mouse and rat cerebral cortex tissues. ICC/IF: HeLa and PC12 cells.
  • General notes

    Ab228477 is the carrier-free version of ab32571. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab228477 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab228477 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 Use at an assay dependent concentration. Detects a band of approximately 116 kDa (predicted molecular weight: 116 kDa).
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Involved in calcium induced regulation of ion channel and activation of the map kinase signaling pathway. May represent an important signaling intermediate between neuropeptide activated receptors or neurotransmitters that increase calcium flux and the downstream signals that regulate neuronal activity. Interacts with the SH2 domain of Grb2. May phosphorylate the voltage-gated potassium channel protein Kv1.2. Its activation is highly correlated with the stimulation of c-Jun N-terminal kinase activity. Involved in osmotic stress-dependent SNCA 'Tyr-125' phosphorylation. In concert with SRC, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. The Tyr-402 phosphorylated form serves as a docking site for SRC and is important for the organization of the osteoclast actin cytoskeleton and attachment sites and for bone resorption.
    • Tissue specificity

      Most abundant in the brain, with highest levels in amygdala and hippocampus. Low levels in kidney. Also expressed in spleen and lymphocytes.
    • Sequence similarities

      Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
      Contains 1 FERM domain.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications

      Phosphorylated on tyrosine residues in response to various stimuli that elevate the intracellular calcium concentration, as well as by PKC activation. Recruitment by nephrocystin to cell matrix adhesions initiates Tyr-402 phosphorylation. In monocytes, adherence to substrata is required for tyrosine phosphorylation and kinase activation. Angiotensin II, thapsigargin and L-alpha-lysophosphatidic acid (LPA) also induce autophosphorylation and increase kinase activity.
    • Cellular localization

      Cytoplasm. Cell membrane. Interaction with nephrocystin induces the membrane-association of the kinase.
    • Information by UniProt
    • Database links

    • Alternative names

      • CADTK antibody
      • CAK-beta antibody
      • CAKB antibody
      • CAKbeta antibody
      • Calcium regulated non receptor proline rich tyrosine kinase antibody
      • Calcium-dependent tyrosine kinase antibody
      • Cell adhesion kinase beta antibody
      • E430023O05Rik antibody
      • EC 2.7.10.2 antibody
      • FADK 2 antibody
      • FADK2 antibody
      • FAK2 antibody
      • FAK2_HUMAN antibody
      • Focal adhesion kinase 2 antibody
      • MGC124628 antibody
      • PKB antibody
      • Proline-rich tyrosine kinase 2 antibody
      • Protein kinase B antibody
      • Protein Tyrosine Kinase 2 Beta antibody
      • Protein-tyrosine kinase 2-beta antibody
      • PTK antibody
      • PTK2B antibody
      • PTK2B protein tyrosine kinase 2 beta antibody
      • PYK2 antibody
      • RAFTK antibody
      • RAFTK2 antibody
      • Related adhesion focal tyrosine kinase antibody
      see all

    Images

    • This WB data was generated using the same anti-PYK2 antibody clone, YE353, in a different buffer formulation (cat# ab32571).

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)              
      Lane 2: PYK2 knockout  HAP1 whole cell lysate (20 µg)
      Lane 3: Jurkat whole cell lysate (20 µg)
      Lane 4: Hu brain whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab32571 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32571 was shown to recognize PYK2 when PYK2 knockout samples were used, along with additional cross-reactive bands. Wild-type and PYK2 knockout samples were subjected to SDS-PAGE.  Ab32571 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PYK2 with purified ab32571 at 1/60. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32571).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue labelling PYK2 with purified ab32571 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32571).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue labelling PYK2 with purified ab32571 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32571).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling PYK2 with purified ab32571 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32571).

    • Immunocytochemistry/Immunofluorescence analysis of PC12 cells labelling PYK2 with unpurified ab32571 at a 1/100 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32571).

    • This IHC data was generated using the same anti-PYK2 antibody clone, YE353, in a different buffer formulation (cat# ab32571).

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human brain tissue labelling PYK2 with unpurified ab32571 at a 1/250 dilution.

    References

    ab228477 has not yet been referenced specifically in any publications.

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