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Rabbit Recombinant Monoclonal Paxillin antibody. Suitable for IP, ELISA, Dot, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Synthetic peptide - Human samples. Cited in 21 publications.

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Images

Immunoprecipitation - Anti-Paxillin antibody [E228] (AB32115), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] (AB32115), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] (AB32115), expandable thumbnail
  • Western blot - Anti-Paxillin antibody [E228] (AB32115), expandable thumbnail
  • Western blot - Anti-Paxillin antibody [E228] (AB32115), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPELISADotWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Expected
Expected
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Expected
Tested
Expected
Expected
Expected
Rat
Expected
Expected
Expected
Tested
Expected
Expected
Expected
Synthetic peptide - Human
Not recommended
Tested
Tested
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/20
Notes

For unpurified use at 1/100

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Expected
Expected

Species
Human, Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
1/1000
Notes

-

Expected
Expected

Species
Human, Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/10000
Notes

-

Species
Rat
Dilution info
1/10000
Notes

-

Species
Human
Dilution info
1/10000
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

For unpurified use at 1/250

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/20
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/50
Notes

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

For unpurified use at 1/100 - 1/250

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). Recruits other proteins such as TRIM15 to focal adhesion.

Alternative names

Paxillin, PXN

Recommended products

Rabbit Recombinant Monoclonal Paxillin antibody. Suitable for IP, ELISA, Dot, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Synthetic peptide - Human samples. Cited in 21 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
E228
Purification technique
Affinity purification Protein A
Specificity

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

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

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 monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Paxillin also known as PXN is a cytoskeletal protein involved in the assembly of focal adhesions. It has a molecular weight of approximately 68 kDa. You will find Paxillin expressed widely in various tissues including the liver colon and lung. Paxillin plays a part in cell motility by interacting with other focal adhesion proteins. It plays a significant mechanical role by serving as a docking site for structural and signaling molecules.

Biological function summary

Paxillin serves key functions in cellular processes such as migration proliferation and survival. It interacts with different proteins including vinculin and talin forming part of a complex at the focal adhesion sites. The phosphorylation state of Paxillin can modulate its interactions and in this way influence the assembly of signaling complexes that control dynamic cellular processes.

Pathways

Paxillin participates in the integrin signaling and MAPK pathways. It operates by linking integrin receptors to the actin cytoskeleton facilitating signal transduction. Paxillin phosphorylation is an important regulatory mechanism within these pathways. In particular its interaction with focal adhesion kinase (FAK) and Src family kinases signifies its role in transmitting signals from the extracellular matrix to the cellular interior which impacts cell behavior and response.

Associated diseases and disorders

Paxillin has links with osteosarcoma and lung cancer. Deregulation of Paxillin expression or its phosphorylation state contributes to oncogenic transformation and tumor progression. In cancer Paxillin associates with altered activity of proteins such as Src and FAK exacerbating disease states. This relationship highlights Paxillin's potential as a biomarker and therapeutic target in treating related malignancies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

  • Immunoprecipitation - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Immunoprecipitation - Anti-Paxillin antibody [E228] (ab32115)

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32115 in HEK-293 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-Paxillin antibody [E228] (ab32115)

    Predicted band size: 65 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] (ab32115)

    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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail
    Image from Beheshti Zavareh R et al. PLoS One. 2012;7(9):e43721. doi: 10.1371/journal.pone.0043721. Epub 2012 Sep 5. Fig 3.; doi:10.1371/journal.pone.0043721; September 5 2012 PLoS ONE 7(9): e43721.

    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] (ab32115)

    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.

  • Western blot - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Western blot - Anti-Paxillin antibody [E228] (ab32115)

    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

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

    Lane 1: HEK-293 (Human embryonic kidney) whole cell lysates at 15 µg

    Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

    Lane 3: C2C12 (Mouse myoblasts myoblast) whole cell lysates at 15 µg

    Lane 4: Rat-1 (Rat embryonic fibroblast) whole cell lysates at 15 µg

    Secondary

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

    Predicted band size: 65 kDa

    Observed band size: 60 kDa, 64 kDa

  • Western blot - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Western blot - Anti-Paxillin antibody [E228] (ab32115)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32115 observed at 65 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab32115 was shown to specifically react with PXN in wild-type A-431 cells as signal was lost in PXN knockout cell line Human PXN (Paxillin) knockout A-431 cell line ab261892 (knockout cell lysate Human PXN (Paxillin) knockout A-431 cell lysate ab261701). Wild-type and PXN knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab32115 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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

    Lane 1: Wild-type A431 whole cell lysate at 20 µg

    Lane 2: PXN knockout A431 whole cell lysate at 20 µg

    Lane 2: Western blot - Human PXN (Paxillin) knockout A-431 cell line (Human PXN (Paxillin) knockout A-431 cell line ab261892)

    Lane 3: U-87 MG whole cell lysate at 20 µg

    Lane 4: PC-3 whole cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 65 kDa

    Observed band size: 65 kDa

    Exposure time: 10s

  • Flow Cytometry (Intracellular) - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Paxillin antibody [E228] (ab32115)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with Purified ab32115 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488, Goat Anti-Rabbit IgG H&L (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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [E228] (ab32115)

    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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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.

  • ELISA - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    ELISA - Anti-Paxillin antibody [E228] (ab32115)

    Direct ELISA antigen dose-response curve using ab32115 at 0-1000 ng/mL. Antigen (human Paxillin phospho Y31 peptide/ unmodified peptide) concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

  • Western blot - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Western blot - Anti-Paxillin antibody [E228] (ab32115)

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

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

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

    Lane 2: HEK-293 (human embryonic kidney) whole cell lysates at 15 µg

    Secondary

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

    Predicted band size: 65 kDa

    Observed band size: 64 kDa

    Exposure time: 3min

  • Dot Blot - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Dot Blot - Anti-Paxillin antibody [E228] (ab32115)

    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. Goat Anti-Rabbit IgG H&L (HRP) 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.

  • Western blot - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Western blot - Anti-Paxillin antibody [E228] (ab32115)

    All lanes: Western blot - 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: 65 kDa

    Observed band size: 64 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [E228] (ab32115), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [E228] (ab32115)

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

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Product protocols

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