Recombinant
RabMAb

Recombinant Anti-PAK1 (phospho T212) antibody [EPR20045] (ab223852)

Overview

  • Product name

    Anti-PAK1 (phospho T212) antibody [EPR20045]
    See all PAK1 primary antibodies
  • Description

    Rabbit monoclonal [EPR20045] to PAK1 (phospho T212)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Dot blot, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human PAK1 aa 200-300 (phospho T212).
    Database link: Q13153

  • Positive control

    • Dot blot: PAK1 (phospho T212) peptide. IHC-P: Human prostate tissue, lung squamous cancer and colon cancer tissue. ICC/IF: HeLa and A549 cells. Flow cytometry: HeLa cells.
  • General notes

    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 ab223852 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
Dot blot 1/1000.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/50.

Target

  • Function

    The activated kinase acts on a variety of targets. Likely to be the GTPase effector that links the Rho-related GTPases to the JNK MAP kinase pathway. Activated by CDC42 and RAC1. Involved in dissolution of stress fibers and reorganization of focal complexes. Involved in regulation of microtubule biogenesis through phosphorylation of TBCB. Activity is inhibited in cells undergoing apoptosis, potentially due to binding of CDC2L1 and CDC2L2.
  • Sequence similarities

    Belongs 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

    Autophosphorylated when activated by CDC42/p21 and RAC1.
  • Cellular localization

    Cytoplasm. Cell junction > focal adhesion. Recruited to focal adhesions upon activation.
  • Information by UniProt
  • Database links

  • Alternative names

    • ADRB2 antibody
    • Alpha PAK antibody
    • Alpha-PAK antibody
    • MGC130000 antibody
    • MGC130001 antibody
    • p21 activated kinase 1 antibody
    • p21 protein (Cdc42/Rac) activated kinase 1 antibody
    • p21-activated kinase 1 antibody
    • p21/Cdc42/Rac1 activated kinase 1 (yeast Ste20 related) antibody
    • p21/Cdc42/Rac1-activated kinase 1 (STE20 homolog, yeast) antibody
    • p65 PAK antibody
    • p65-PAK antibody
    • P68-PAK antibody
    • PAK alpha antibody
    • PAK-1 antibody
    • Pak1 antibody
    • PAK1_HUMAN antibody
    • Paka antibody
    • PAKalpha antibody
    • Protein kinase MUK2 antibody
    • Rac/p21-activated kinase antibody
    • Serine/threonine-protein kinase PAK 1 antibody
    • STE20 homolog yeast antibody
    see all

Images

  • Dot blot analysis of PAK1 (phospho T212) peptide using ab223852 1/1,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (ab97051), was used as secondary antibody at 1/100,000 dilution.

    Lane 1: PAK1 (phospho T212) peptide.

    Lane 2: PAK1 non-phospho peptide.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling PAK1 (phospho T212) with ab223852 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing positive staining on HeLa cells in M phase. The signal decreased after treatment with Lambda Protein Phosphatase 31℃ for 2 hours (treated).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    The negative control: PBS followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human lung squamous cancer tissue labeling PAK1 (phospho T212) with ab223852 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on tumor cells of human lung squamous cancer (PMID: 19876919; PMID: 12176334) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

     

  • Flow cytometric analysis of 80% methanol-fixed, 0.1% Tween-20 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling PAK1 (phospho T212) with ab223852 at 1/50 (right panel) compared with a Rabbit monoclonal IgG (ab172730) Isotype control (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2,000 dilution was used as the secondary antibody. Unlabeled control cells without incubation with primary antibody and secondary antibody (Blue).

    Cells were pretreated with 20 μg/ml RNase A for 30 minutes to eliminate the non-specific binding between RNA and PI (Propidium iodide). PAK1 (phospho T212) is highly expressed in mitotic cells (PMID: 12176334).

     

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PAK1 (phospho T212) with ab223852 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on tumor cells of human colon cancer (PMID: 19876919; PMID: 12176334) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human prostate tissue labeling PAK1 (phospho T212) with ab223852 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadic nuclear staining on human prostate (PMID: 19876919; PMID: 12176334) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling PAK1 (phospho T212) with ab223852 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing positive staining on HeLa cells in M phase. The signal decreased after treatment with Lambda Protein Phosphatase 31℃ for 2 hours (treated).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    The negative control: PBS followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution.

References

ab223852 has not yet been referenced specifically in any publications.

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