Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1473] to CDKN2A/p16INK4a - BSA and Azide free
  • Suitable for: ICC, WB, IHC-P, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free
    See all CDKN2A/p16INK4a primary antibodies

  • Description

    Rabbit monoclonal [EPR1473] to CDKN2A/p16INK4a - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • HeLa, 293T, and Saos-2 cell lysates, Human cervical carcinoma tissue, HeLa cells

  • General notes

    Ab186932 is the carrier-free version of ab108349. 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

    ab186932 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932) Image from Shan M et al.,; PLoS One. 2013;8(10):e76408. Fig 1.; doi: 10.1371/journal.pone.0076408. eCollection 2013.

    Formalin-fixed, paraffin-embedded human normal breast, luminal-A DCIS (ductal carcinoma in situ) and triple negative breast cancer tissues stained for CDKN2A/p16INK4a using ab108349 in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

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

  • Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    ab108349 (purified) at 1/30 immunoprecipitating CDKN2A/p16INK4a in HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

  • Flow Cytometry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Flow Cytometry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Flow Cytometry analysis of HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling CDKN2A/p16INK4a with purified ab108349 at 1/270 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

  • Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Clone EPR1473 (ab186932) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4a antibody [EPR1473] (PE). Please refer to ab209579 for protocol details.

    ab209579 staining CDKN2A/p16INK4a in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209579 at 1/500 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Clone EPR1473 (ab186932) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4a antibody [EPR1473] (Alexa Fluor® 488). Please refer to ab192053 for protocol details.

    ab192053 staining CDKN2A/p16INK4a in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab192053 at a working dilution of 1 in 50 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Clone EPR1473 (ab186932) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4a antibody [EPR1473] (Alexa Fluor® 647). Please refer to ab192054 for protocol details.

    ab192054 staining CDKN2A/p16INK4a in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab192054 at a working dilutions of 1 in 100 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling CDKN2A/p16INK4a with purified ab108349 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

    Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling CDKN2A/p16INK4a with unpurified ab108349 at a dilution of 1/250.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

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

  • Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Immunocytochemistry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling CDKN2A/p16INK4a with unpurified ab108349 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

  • Flow Cytometry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Flow Cytometry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Overlay histogram showing HEK-293 (Human epithelial cell line from embryonic kidney) cells stained with unpurified ab108349 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108349, 1/100) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK-293 cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108349).

  • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932) + HEK293 (human embryonic kidney) whole cell lysate at 10 µg

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

    Predicted band size: 17 kDa


    Exposure time: 3 minutes


    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)
    Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

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