Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y113] to Paxillin - BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Human

Overview

  • Product name

    Anti-Paxillin antibody [Y113] - BSA and Azide free
    See all Paxillin primary antibodies

  • Description

    Rabbit monoclonal [Y113] to Paxillin - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Mouse
    WB
    Human
    See all applications and species data

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK0293 and A431 cell lysates.

  • General notes

    Ab216652 is the carrier-free version of ab32084. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab216652 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Western blot - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    All lanes : Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PXN knockout HeLa cell lysate
    Lane 3 : HEK-293 cell lysate
    Lane 4 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab32084).

    Lanes 1-4: Merged signal (red and green). Green - ab32084 observed at 68 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab32084 Anti-Paxillin antibody [Y113] was shown to specifically react with Paxillin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264767 (knockout cell lysate ab257044) was used. Wild-type and Paxillin knockout samples were subjected to SDS-PAGE. ab32084 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652) Image from Fuhrmann A et al. PLoS One. 2014;9(7):e102424. Fig 3.; doi: 10.1371/journal.pone.0102424.

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • Flow Cytometry - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Flow Cytometry - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652) This image is courtesy of an Abreview submitted by Shaohua Li.

    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

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)

    ab32084 showing positive staining in Normal ovary tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)

    ab32084 showing positive staining in Ovarian carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652) This image is courtesy of an anonymous Abreview.
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • OI-RD Scanning - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    OI-RD Scanning - Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

  • Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)
    Anti-Paxillin antibody [Y113] - BSA and Azide free (ab216652)

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