Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9518] to PODXL - BSA and Azide free
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PODXL antibody [EPR9518] - BSA and Azide free
    See all PODXL primary antibodies

  • Description

    Rabbit monoclonal [EPR9518] to PODXL - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human PODXL aa 300-500. The exact sequence is proprietary.
    Database link: O00592

  • Positive control

    • WB: HCT116, HAP1, Raji and HeLa cell lysates. Human fetal kidney tissue lysate. IHC-P: Human kidney tissue. Human hepatocellular carcinoma, breast carcinoma and endometrial carcinoma tissue. Human glioma. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    ab269888 is the carrier-free version of ab150358. 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

    ab269888 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    All lanes : Anti-PODXL antibody [EPR9518] (ab150358) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PODXL knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 160 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab150358).

      Lanes 1- 2: Merged signal (red and green). Green - ab150358 observed at 160 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab150358 was shown to react with PODXL in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264984 (knockout cell lysate ab257210) was used. Wild-type HeLa and PODXL knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab150358 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 Dilution and a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded human breast adenocarcinoma tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    All lanes : Anti-PODXL antibody [EPR9518] (ab150358) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : PODXL knockout HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab150358).

    Lanes 1- 2: Merged signal (red and green). Green - ab150358 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab150358 was shown to react with PODXL in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266887 (knockout cell lysate ab257211) was used. Wild-type HCT116 and PODXL knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab150358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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.

  • Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Western blot - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    All lanes : Anti-PODXL antibody [EPR9518] (ab150358) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PODXL knockout HAP1 whole cell lysate
    Lane 3 : Raji whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 58 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab150358 observed at 160 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab150358 was shown to specifically react with PODXL in wild-type cells as signal was lost in PODXL knockout cells. Wild-type and PODXL knockout samples were subjected to SDS-PAGE. Ab150358 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Flow Cytometry - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Flow Cytometry - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling PODXL with purified ab150358 at 1/250 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells 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 (ab150358).

  • Immunocytochemistry/ Immunofluorescence - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunocytochemistry/ Immunofluorescence - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labeling PODXL with purified ab150358 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei couterstained with DAPI (blue).

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded human glioma tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded normal human kidney tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded human skeletal muscle tissue using unpurified ab150358 showing negative staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded human endometrial carcinoma tissue using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded human hepatocellular carcinoma vessels using unpurified ab150358 showing positive staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemical analysis of paraffin embedded human kidney tissue labeling PODXL with unpurified ab150358 antibody at a dilution of 1/100. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling PODXL with purified ab150358 at 1/1000 dilution (0.44 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) is the secondary antibody.

    PBS instead of the primary antibody was used as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150358).

  • Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)
    Anti-PODXL antibody [EPR9518] - BSA and Azide free (ab269888)

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