Anti-PYK2 (phospho Y402) antibody (ab4800)

Submit a review Q&A (3)References (8)
Western blot - Anti-PYK2 (phospho Y402) antibody (ab4800)
  • Immunocytochemistry/ Immunofluorescence - Anti-PYK2 (phospho Y402) antibody (ab4800)

Key features and details

  • Rabbit polyclonal to PYK2 (phospho Y402)
  • Suitable for: ICC/IF, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-PYK2 (phospho Y402) antibody
    See all PYK2 primary antibodies

  • Description

    Rabbit polyclonal to PYK2 (phospho Y402)

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide (Human) derived from the region of human Pyk 2 that contains tyrosine 402. The sequence is conserved in human and rat.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab4800 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Mouse
WB
Human
Application Abreviews Notes
ICC/IF
WB
Notes
  • Application notes
    IHC-Fr: Use at an assay dependent dilution.
    WB: 1/1000. Predicted molecular weight: 116 kDa. Can be blocked with PYK2 peptide - phospho Y402 (phospho and non-phospho pair) or CEF whole cell lysate, positive / negative for PYK 2 (pair).
    ICC/IF: Use at an assay dependent dilution from (PubMed:17325207).
    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.

    • 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

      • Western blot - Anti-PYK2 (phospho Y402) antibody (ab4800)
        Western blot - Anti-PYK2 (phospho Y402) antibody (ab4800)
        Peptide Competition: Extracts prepared from CEF cells expressing Pyk2 and plated on fibronectin were resolved by SDS-PAGE on a 10% Tris glycine gel. The proteins were then transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.50 µg/mL ab4800 antibody, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to phosphopeptide immunogen (2), a generic phosphotyrosine containing peptide (3), the phosphopeptide corresponding to tyrosine 397 on FAK (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, therefore demonstrating the specificity of ab4800 antibody for this phosphorylated residue. Peptide Competition: Extracts p
      • Immunocytochemistry/ Immunofluorescence - Anti-PYK2 (phospho Y402) antibody (ab4800)
        Immunocytochemistry/ Immunofluorescence - Anti-PYK2 (phospho Y402) antibody (ab4800)
        Figure shows Pyk2 staining: Normal mouse mammary (NmuMG) cells were treated with TGFbeta and then grown to confluence, fixed and permeabilized. ab4800 was identified using a selective PSSA followed by incubation with a Cy3-conjugated anti-rabbit secondary antibody (red). F actin was visualized with Oregon Green-conjugated phalloiden (green) and paxillin staining is indicated in blue. The merged image indicates colocalization (purple) of ab4800, F-actin and paxillin in focal adhesion structures.

      References (8)

      Publishing research using ab4800? Please let us know so that we can cite the reference in this datasheet.

      ab4800 has been referenced in 8 publications.

      • Sai C  et al. Cyclin Dependent Kinase 1 (CDK1) Activates Cardiac Fibroblasts via Directly Phosphorylating Paxillin at Ser244. Int Heart J 60:374-383 (2019). PubMed: 30745530
      • Wang Q  et al. Small molecule FAK activator promotes human intestinal epithelial monolayer wound closure and mouse ulcer healing. Sci Rep 9:14669 (2019). PubMed: 31604999
      • Tiede S  et al. The FAK inhibitor BI 853520 exerts anti-tumor effects in breast cancer. Oncogenesis 7:73 (2018). PubMed: 30237500
      • Haidari M  et al. Disruption of endothelial adherens junctions by high glucose is mediated by protein kinase C-ß-dependent vascular endothelial cadherin tyrosine phosphorylation. Cardiovasc Diabetol 13:112 (2014). Human . PubMed: 25027642
      • Haidari M  et al. Atorvastatin preserves the integrity of endothelial adherens junctions by inhibiting vascular endothelial cadherin tyrosine phosphorylation. Exp Cell Res 318:1673-84 (2012). WB ; Human . PubMed: 22659162
      • Xie H  et al. Silencing or Amplification of Endocannabinoid Signaling in Blastocysts via CB1 Compromises Trophoblast Cell Migration. J Biol Chem 287:32288-97 (2012). WB . PubMed: 22833670
      • Suo L  et al. Protocadherin clusters and cell adhesion kinase regulate dendrite complexity through Rho GTPase. J Mol Cell Biol : (2012). PubMed: 22730554
      • Schober M  et al. Focal adhesion kinase modulates tension signaling to control actin and focal adhesion dynamics. J Cell Biol 176:667-80 (2007). PubMed: 17325207

      Customer reviews and Q&As

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      1-3 of 3 Abreviews or Q&A

      Question

      Phone call requesting information on if the Anti-PYK2 (phospho Y402) antibody (ab4800) is likely to cross react with other phospho-proteins.

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      Answer

      Thank you for contacting us and your interest in our products. I am sorry for the delay in getting back to you.

      As we discussed over the phone, I have looked into how the Anti-PYK2 (phospho Y402) antibodies ab4800 and ab131543 have been characterized for their potential cross-reactivity with other phospho-proteins.

      No specific experiments have been carried out to ascertain the cross-reactivity of either of these antibodies. However, the western results are based on total cell lysates which contains thousands of other proteins many of which are phosphorylated. As the Western blot images for these antibodies show, only one band was detected in the molecular range from 37 kDa up to 250 kDa, there is no other visible band.

      In addition, with ab4800 the target band was competed out with the phosphorylated immunogen peptide indicating the band we see is the targe Pyk2 pY402. This supports at least in the cells (primary CDF cells, and SDF-1a treated Reh cells) we studied we did not see any cross reactivity with other proteins (phosphorylated or not phosphorylated).

      From blast search of the immunogens used, the area does not seem to share homology with any other protein (that have been added to this databank) suggesting there should not be any cross-reactivity observed.

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      Question

      Hi,   I have technical questions regarding 2 of your products: PYK2 protein (Active), cat. no ab60878, and PYK2 protein (His tag), cat. no ab42622.   I need to know the phosphorylation site of these 2 proteins. More specifically, I need to know if these proteins are phosphorylated at Tyrosine 402.  

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      Answer

      Thank you for contacting us. I do want to apologize for the tardiness of this response.

      I have been in contact with the originator labs for ab42622 and ab60878 PYK2 proteins, in regards to whether these products are phosphorylated at Y402. Unfortunately neither group was able to confirm whether Y402 is phosphorylated at this site and each recommended that phophorylation state may be determined by performing a western blot with a phospho specific antibody such as ab4800 (https://www.abcam.com/PYK2-phospho-Y402-antibody-ab4800.html).

      I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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      Question

      I am interested in ab4800, the Pyk2 phospho Y402 antibody. We would like to use it on mouse pituitary glands. Can you tell me if this antibody works with paraffin-embedded sections? If not under what conditions might it work in tissues rather than cells (your references on the web site are all cells)?

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      Answer

      Thank you for your enquiry. Ab4800 was tested in IHC cryostat sections in the following reference: Melendez J. et al (2002) J. Biol. Chem. 277(47):45203-45210. The authors prepared mouse heart sections fixed in 4% paraformaldehyde/PBS overnight at 4 degrees C. The next day, the hearts were subjected to progressive sucrose gradient from 10%, 20%, 30% at 4 degrees C for 1 hour at each step. Embedded in Tisse-Tek, frozen in melting isopentane floated above liquid nitrogen, and frozen blocks were stored at -80 degrees C Section 7 microns thick were mounted on Superfrost slides (Fisher Scientific Co), allowed to dry, and then stored at 4 degrees C until used within 1 week. For labeling, primary antibodies were applied to the sections and left overnight. The next day, the sections were washed, secondary antibodies applied to the sections for 1 hour, washed again, and then mounted in Vectashield (Vecyor Laboratories). Primary antibodies were detected by fluorescent secondary goat anti-rabbit IgG. We recommend using the antibody at ~1:50 dilution to 1:100 dilution. If you have any more questions, please contact us again.

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