Overview

  • Product name
    Anti-Paxillin antibody [E228]
    See all Paxillin primary antibodies
  • Description
    Rabbit monoclonal [E228] to Paxillin
  • Host species
    Rabbit
  • Specificity
    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-P, Flow Cyt, IP, Dot blot, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Paxillin aa 1-100 (phospho Y31). The exact sequence is proprietary.
    Database link: P49023
    (Peptide available as ab179557)

  • Positive control
    • WB: NIH 3T3, HEK-293, C2C12, and Rat-1 cell lysate. ICC/IF: HeLa cells. IHC-P: Human colon carcinoma, Human breast carcinoma. FC: HeLa cells IP: HEK-293 cell lysate
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32115 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 1/10000. Predicted molecular weight: 64 kDa.
ICC/IF 1/100.

For unpurified use at 1/250

IHC-P 1/50.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250

Flow Cyt 1/20.
IP 1/20.

For unpurified use at 1/100

Dot blot 1/1000.
ELISA Use at an assay dependent concentration.

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

  • All lanes : Anti-Paxillin antibody [E228] (ab32115) at 1/1000 dilution (purified)

    Lane 1 : HEK-293 (Human embryonic kidney) whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 3 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
    Lane 4 : Rat-1 (Rat embryonic fibroblast) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 64 kDa
    Observed band size: 60,64 kDa
    why is the actual band size different from the predicted?



    Based on the immunogen sequence blast, this antibody recognizes alpha, beta and gamma isoforms. The molecular weight observed is consistent with what has been described in the literature PMID: 9712867 and 20388733

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling Paxillin with Purified ab32115 at 1:50 dilution (2.34 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with Purified ab32115 at 1:100 dilution (1.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • ab32115 (purified) at 1:20 dilution (0.5µg) immunoprecipitating Paxillin in HEK-293 whole cell lysate.
    Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab32115 & HEK-293 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32115 in HEK-293 whole cell lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with Purified ab32115 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Paxillin with ab32115 at 1/100 (1 μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/ml) was used as the secondary antibody. The cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclei counterstained with DAPI (blue). 

    Confocal image showing membranous staining on HeLa cells.

  • All lanes : Anti-Paxillin antibody [E228] (ab32115) at 1/1000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : HEK-293 (human embryonic kidney) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 64 kDa
    Observed band size: 64 kDa


    Exposure time: 3 minutes


    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  • Immunofluorescence analysis of HeLa cells, staining Paxillin with ab32115.

    Cells on the right were treated with MGAT1 shRNA. Cells were fixed with 2% paraformaldehyde, permeabilized using 0.2% Triton-X-100 and blocked by 5% BSA for 1 hour. Cells were incubated with primary antibody (1/400) overnight at 4°C. A FITC-conjugated donkey anti-rabbit IgG (1/500) was used as the secondary antibody.

  • All lanes : Anti-Paxillin antibody [E228] (ab32115) at 1/10000 dilution

    Lane 1 : untreated NIH 3T3 cell lysate
    Lane 2 : PDGF treated NIH 3T3 cell lysate

    Predicted band size: 64 kDa
    Observed band size: 64 kDa

  • Direct ELISA antigen dose-response curve using ab32115 at 0-1000 ng/mL. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

  • Dot blot analysis of Paxillin (pY31) peptide (Lane 1) and Paxillin non-phospho peptide (Lane 2) labelling Paxillin with ab32115 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemical analysis of Paxillin expression in paraffin-embedded human colon carcinoma using 1/100 ab32115.

References

This product has been referenced in:
  • Tsai CS  et al. Thrombomodulin regulates monocye differentiation via PKCd and ERK1/2 pathway in vitro and in atherosclerotic artery. Sci Rep 6:38421 (2016). WB ; Human . Read more (PubMed: 27910925) »
  • Sausgruber N  et al. Tyrosine phosphatase SHP2 increases cell motility in triple-negative breast cancer through the activation of SRC-family kinases. Oncogene N/A:N/A (2014). WB . Read more (PubMed: 24931162) »
See all 5 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab32115.
Please use the links above to contact us or submit feedback about this product.

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

Sign up