Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17878] to CDKN2A/p14ARF - BSA and Azide free
  • Suitable for: Flow Cyt, IP, ICC/IF, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free
    See all CDKN2A/p14ARF primary antibodies

  • Description

    Rabbit monoclonal [EPR17878] to CDKN2A/p14ARF - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HeLa whole cell lysate transfected with CDKN2A/p14ARF with His-tag; PC-3 and HEK-293 whole cell lysates. ICC/IF: HeLa cells transfected with CDKN2A/p14ARF. Flow Cyt: HeLa cells transfected with CDKN2A/p14ARF-GFP. IP: CDKN2A/p14ARF transfected HeLa whole cell lysate.

  • General notes

    Ab224273 is the carrier-free version of ab185620. 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

    ab224273 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

  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF or empty vector, labeling CDKN2A/p14ARF with ab185620 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab185620 at 1/250 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185620).

  • Flow Cytometry - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    Flow Cytometry - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF-GFP labeling CDKN2A/p14ARF with ab185620 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (PE) at 1/150 dilution 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 (ab185620).

  • Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

    CDKN2A/p14ARF was immunoprecipitated from 1mg of CDKN2A/p14ARF transfected HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab185620 at 1/80 dilution.

    Western blot was performed from the immunoprecipitate using ab185620 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: CDKN2A/p14ARF transfected HeLa whole cell lysate, 10µg (Input).

    Lane 2: ab185620 IP in CDKN2A/p14ARF transfected HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal[EPR25A] - Isotype Control (ab172730) instead of ab185620 in CDKN2A/p14ARF transfected HeLa whole cell lysate.

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

    Exposure time: 1 second.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185620).

  • Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/1000 dilution

    Lane 1 : Wild type HeLa whole cell lysate
    Lane 2 : CDKN2A knockout HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 14 kDa



    Lanes 1 - 2: Merged signal (red and green). Green - ab185620 observed at 14 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab185620 was shown to specifically react with CDKN2A/p14ARF in wild-type HeLa cells as signal was lost in CDKN2A knockout cells. Wild-type and CDKN2A knockout samples were subjected to SDS-PAGE. Ab185620 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 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 (ab185620).

  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

    This ICC data was generated using the same anti-CDKN2A/p14ARF antibody clone [EPR17878] in a different buffer formulation (cat# ab185620).

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF or empty vector, labeling CDKN2A/p14ARF with ab185620 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab185620 at 1/250 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)
    Anti-CDKN2A/p14ARF antibody [EPR17878] - BSA and Azide free (ab224273)

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