Overview

  • Product nameAnti-PAK2 antibody [EP796Y]
    See all PAK2 primary antibodies
  • Description
    Rabbit monoclonal [EP796Y] to PAK2
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues near the N terminus of human PAK2.

  • Positive control
    • WB: HeLa cytoplasmic lysate IHC-P; Human breast carcinoma tissue IF/ICC: T47D cell line.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

Applications

Our Abpromise guarantee covers the use of ab76293 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
ICC/IF 1/100 - 1/250.
WB 1/1000 - 1/2000. Predicted molecular weight: 58 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use of HRP conjugated or polymerized HRP secondary antibody is recommended. Stronger signals have been found using the polymerized HRP secondary.

Flow Cyt 1/100.

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

  • Application notesIs unsuitable for IP.
  • Target

    • FunctionThe activated kinase acts on a variety of targets. Phosphorylates ribosomal protein S6, histone H4 and myelin basic protein. Full length PAK 2 stimulates cell survival and cell growth. The process is, at least in part, mediated by phosphorylation and inhibition of pro-apoptotic BAD. Caspase-activated PAK-2p34 is involved in cell death response, probably involving the JNK signaling pathway. Cleaved PAK-2p34 seems to have a higher activity than the CDC42-activated form.
    • Tissue specificityUbiquitously expressed. Higher levels seen in skeletal muscle, ovary, thymus and spleen.
    • Sequence similaritiesBelongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.
      Contains 1 CRIB domain.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications
      Full length PAK 2 is autophosphorylated when activated by CDC42/p21. Following cleavage, both peptides, PAK-2p27 and PAK-2p34, become highly autophosphorylated, with PAK-2p27 being phosphorylated on serine and PAK-2p34 on threonine residues, respectively. Autophosphorylation of PAK-2p27 can occur in the absence of any effectors and is dependent on phosphorylation of Thr-402, because PAK-2p27 is acting as an exogenous substrate.
      During apoptosis proteolytically cleaved by caspase-3 or caspase-3-like proteases to yield active PAK-2p34.
      Ubiquitinated, leading to its proteasomal degradation.
      PAK-2p34 is myristoylated.
    • Cellular localizationCytoplasm and Nucleus. Cytoplasm > perinuclear region. Membrane. Interaction with ARHGAP10 probably changes PAK-2p34 location to cytoplasmic perinuclear region. Myristoylation changes PAK-2p34 location to the membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • C-t-PAK2 antibody
      • CB422 antibody
      • EC 2.7.11.1 antibody
      • Gamma PAK antibody
      • Gamma-PAK antibody
      • hPAK65 antibody
      • Kinase antibody
      • p21 (CDKN1A) activated kinase 2 antibody
      • p21 (CDKN1A)-activated kinase 2a antibody
      • p21 activated kinase 2 antibody
      • p21 protein (Cdc42/Rac)-activated kinase 2 antibody
      • p21 protein Cdc42 Rac activated kinase 2 antibody
      • p21-activated kinase 2 antibody
      • p21-activated kinase, 65-KD antibody
      • p21-activated protein kinase I antibody
      • p21CDKN1A activated kinase 2 antibody
      • p27 antibody
      • p34 antibody
      • p58 antibody
      • p65PAK antibody
      • PAK 2 antibody
      • PAK-2 antibody
      • PAK-2p34 antibody
      • Pak2 antibody
      • PAK2_HUMAN antibody
      • PAK65 antibody
      • PAKgamma antibody
      • S6 H4 kinase antibody
      • S6/H4 kinase antibody
      • Serine threonine protein kinase PAK 2 antibody
      • Serine/threonine protein kinase PAK 2 antibody
      see all

    Anti-PAK2 antibody [EP796Y] images



    • Predicted band size : 58 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PAK2 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Human lymph node tissue lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab76293 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab76293 was shown to specifically react with PAK2 when PAK2 knockout samples were used. Wild-type and PAK2 knockout samples were subjected to SDS-PAGE. ab76293 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • ICC/IF image of ab76293 stained T47D cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76293, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Overlay histogram showing HeLa cells stained with ab76293 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76293, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • ab76293, at a 1/100 dilution, staining PAK2 in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.
    • Anti-PAK2 antibody [EP796Y] (ab76293) at 1/2000 dilution + HeLa cell lysate at 10 µg

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 58 kDa
      Observed band size : 61 kDa (why is the actual band size different from the predicted?)

    References for Anti-PAK2 antibody [EP796Y] (ab76293)

    This product has been referenced in:
    • Jiang N  et al. In vivo quantitative phosphoproteomic profiling identifies novel regulators of castration-resistant prostate cancer growth. Oncogene N/A:N/A (2014). Mouse . Read more (PubMed: 25065596) »
    • Chen G  et al. TGFß receptor I transactivation mediates stretch-induced Pak1 activation and CTGF upregulation in mesangial cells. J Cell Sci 126:3697-712 (2013). Read more (PubMed: 23781022) »

    See all 3 Publications for this product

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