Recombinant
RabMAb

Anti-Paxillin antibody [Y113] (ab32084)

Overview

  • Product name
    Anti-Paxillin antibody [Y113]
    See all Paxillin primary antibodies
  • Description
    Rabbit monoclonal [Y113] to Paxillin
  • Host species
    Rabbit
  • Specificity
    ab32084 recognises Paxillin alpha, beta and gamma isoforms.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human
  • Immunogen

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

  • Positive control
    • WB: HeLa cell lysate; RAW264.7 whole cell lysate. IHC-P: Human breast carcinoma; Normal ovary tissue; Ovarian carcinoma tissue; Transitional cell carcinoma of kidney tissue. ICC/IF: 3T3 fibroblasts; Mouse embryonic fibroblasts; MEF1 cells treated with (S)-(-)-Blebbistatin; bovine kidney cells. Flow Cyt: 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 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.
Flow Cyt Use at an assay dependent concentration.
IP 1/200.

Target

  • Function
    Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion).
  • Sequence similarities
    Belongs 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 localization
    Cytoplasm > 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

Images

  • Shear-induced Cell Remodeling.

    3T3 fibroblasts are shown under the indicated cation and shear conditions. The shear direction in each image is indicated by a white arrow. Images show paxillin in green (ab32084), the actin cytoskeleton in red, and the nucleus (DNA) in blue. The approximate pre-shear cell area is indicated by white dashed lines as determined from the focal adhesions that remained on the substrate, which are indicated by open arrowheads. The bottom left image was contrast-enhanced 2-fold to better visualize the focal adhesions that remained on the substrate. Inset images are shown from regions outlined in white.

  • 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 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.
  • ab32084 showing positive staining in Ovarian carcinoma tissue.

  • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution ( 10ug/ml) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077) )(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

  • 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.
  • 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



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

    See Abreview

  • 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

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Charrier EE  et al. Control of cell morphology and differentiation by substrates with independently tunable elasticity and viscous dissipation. Nat Commun 9:449 (2018). Read more (PubMed: 29386514) »
  • Lai H  et al. FAK-ERK activation in cell/matrix adhesion induced by the loss of apolipoprotein E stimulates the malignant progression of ovarian cancer. J Exp Clin Cancer Res 37:32 (2018). Read more (PubMed: 29458390) »

See all 54 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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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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