Overview

  • Product nameAnti-Paxillin antibody [Y113]
    See all Paxillin primary antibodies
  • Description
    Rabbit monoclonal [Y113] to Paxillin
  • Specificityab32084 recognises Paxillin alpha, beta and gamma isoforms.
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Paxillin aa 1-100 (N terminal).

  • Positive control
    • HeLa cell lysate, human breast carcinoma.
  • 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.

     

    Alternative versions available:

    Anti-Paxillin antibody (HRP) [Y113] (ab197612)

Properties

Applications

Our Abpromise guarantee covers the use of ab32084 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 1/5000 - 1/10000. Predicted molecular weight: 68 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250.
IP 1/200.
  • Application notesIs unsuitable for Flow Cyt.
  • Target

    • FunctionCytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion).
    • Sequence similaritiesBelongs to the paxillin family.
      Contains 4 LIM zinc-binding domains.
    • Post-translational
      modifications
      Phosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens.
    • Cellular localizationCytoplasm > cytoskeleton. Cell junction > focal adhesion.
    • Information by UniProt
    • Database links
    • Alternative names
      • FLJ16691 antibody
      • FLJ23042 antibody
      • Paired box protein Pax 1 antibody
      • PAX 1 antibody
      • PAX1 antibody
      • PAXI_HUMAN antibody
      • Paxillin alpha antibody
      • Paxillin antibody
      • PXN antibody
      • PXN protein antibody
      see all

    Anti-Paxillin antibody [Y113] images

    • ab32084 staining paxillin in MEF1 cells treated with (S)-(-)-Blebbistatin (ab120491), by ICC/IF. Decreased membrane expression of paxillin correlates with increased concentration of (S)-(-)-Blebbistatin , as described in literature.
      The cells were incubated at 37°C for 2h in media containing different concentrations of ab120491 ( (S)-(-)-Blebbistatin ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32084 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
    • Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

      Sample: mouse embryonic fibroblasts

      Preparation:

      Fix in 3% PFA in PBS for 30 min at RT

      Primary antibody: Rabbit anti-paxillin Y113 (ab32084), 1:100

      Secondary antibody: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200

      Rhodamine-phalloidin, 1:100

      Nuclei were counterstained with DAPI

       

    • Anti-Paxillin antibody [Y113] (ab32084) at 1/10000 dilution + HeLa cell lysate.

      Predicted band size : 68 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)
    • ab32084 showing positive staining in Normal ovary tissue.

    • Anti-Paxillin antibody [Y113] (ab32084) at 1/5000 dilution + Mouse RAW264.7 whole cell lysate at 20 µg

      Secondary
      An HRP Donkey anti-rabbit IgG polyclonal at 1/10000 dilution

      Predicted band size : 68 kDa

      This image is courtesy of an Abreview submitted by Dr. Alban Gaultier

      Blocking step: 5% Milk for 1 hour at 20°C.\
      Gel: SDS-PAGE

      See Abreview

    • ab32084 showing positive staining in Ovarian carcinoma tissue.

    • ab32084 showing positive staining in Transitional cell carcinoma of kidney tissue.

    • Immunofluorescence analysis of bovine kidney cells, staining Paxillin with ab32084.

      Cells were fixed with paraformaldehyde, permeabilized with 1% Triton X-100 and blocked with 5% BSA for 1 hour. Samples were incubated with primary antibody (1/2500 in 5% BSA) for 1 hour at 25°C. An undiluted AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

      See Abreview

    • ab32084 staining paxillin in MEF1 cells treated with (+/-)-blebbistatin (ab120425), by ICC/IF. Decreased membrane expression of paxillin correlates with increased concentration of (+/-)-blebbistatin, as described in literature.
      The cells were incubated at 37°C for 1h in media containing different concentrations of ab120425 ((+/-)-blebbistatin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32084 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References for Anti-Paxillin antibody [Y113] (ab32084)

    This product has been referenced in:
    • Izumi D  et al. CXCL12/CXCR4 activation by cancer-associated fibroblasts promotes integrin ß1 clustering and invasiveness in gastric cancer. Int J Cancer 138:1207-19 (2016). ICC/IF ; Human . Read more (PubMed: 26414794) »
    • Yariswamy M  et al. Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction. J Biol Chem 291:19425-36 (2016). Read more (PubMed: 27466370) »

    See all 29 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Mouse Cell (RAW264.7)
    Specification RAW264.7
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.1% Triton + 2% BSA in PBS
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C
    Username

    Dr. Mahesh Shivananjappa

    Verified customer

    Submitted Apr 16 2013

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HBMEC)
    Permeabilization Yes - 0.1% Triton in PBS
    Specification HBMEC
    Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Nov 19 2015

    Application Western blot
    Sample Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions Reduced Denaturing
    Loading amount 20 µg
    Specification HeLa
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Aug 24 2015

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Sample Human Cell (HeLa cell)
    Specification HeLa cell
    Permeabilization Yes - 1% Triton X-100 in PBS
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 24 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (MDAMB435S)
    Specification MDAMB435S
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.1% Triton in PBS
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 28 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Rat Cell lysate - whole cell (rat benign mammary cell line)
    Loading amount 20 µg
    Specification rat benign mammary cell line
    Gel Running Conditions Reduced Denaturing (12.5%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 04 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Dog Cell lysate - whole cell (MDCK)
    Loading amount 10 µg
    Specification MDCK
    Gel Running Conditions Reduced Denaturing (10% tris-glycine gel)
    Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 08 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (HL60)
    Loading amount 10 µg
    Specification HL60
    Gel Running Conditions Reduced Denaturing (10% tris-glycine)
    Blocking step Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 02 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Cow Cell (Mardin-Darby bovine kidney cells)
    Specification Mardin-Darby bovine kidney cells
    Fixative Paraformaldehyde
    Permeabilization Yes - 1% Triton-X-100 (20 min)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Abcam user community

    Verified customer

    Submitted Oct 16 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Cow Cell lysate - whole cell (MDBK cells)
    Loading amount 10 µg
    Specification MDBK cells
    Gel Running Conditions Reduced Denaturing (10 % Tris-Glycine gel)
    Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 16 2012

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"