Recombinant
RabMAb

Anti-CDKN2A/p16INK4a antibody [EPR1473] (ab108349)

Overview

  • Product name
    Anti-CDKN2A/p16INK4a antibody [EPR1473]
    See all CDKN2A/p16INK4a primary antibodies
  • Description
    Rabbit monoclonal [EPR1473] to CDKN2A/p16INK4a
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CDKN2A/p16INK4a aa 100 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P42771

  • Positive control
    • WB: HeLa, HEK-293, HEK-293T, and Saos-2 cell lysates. IHC-P: Human cervical carcinoma tissue. ICC/IF: HeLa cells. Flow Cytometry: HEK-293 and HeLa cells. IP: HeLa cell lysate.
  • General notes

    A trial size is available to purchase for this antibody.

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108349 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/2000. Predicted molecular weight: 17 kDa.

For unpurified use at 1/1000 - 1/10000.

IP 1/30.

For unpurified use at 1/10 - 1/100.

IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

Flow Cyt 1/270.

For unpurified use at 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/270.

For unpurified use at 1/100 - 1/250.

Target

  • Cellular localization
    Cytoplasmic and Nuclear
  • Database links
  • Form
    There are 4 isoforms produced by alternative splicing. Isoform 1 also known as: p16INK4a; Isoform 3 also known as: p12; Isoform 4 also known as: p14ARF; p19ARF; ARF.
  • Alternative names
    • CCM2 antibody
    • CDK4 inhibitor p16 INK4 antibody
    • CDK4I antibody
    • CDKN2 antibody
    • CDKN2A antibody
    • Cell cycle negative regulator beta antibody
    • CMM2 antibody
    • Cyclin dependent kinase 4 inhibitor A antibody
    • Cyclin dependent kinase inhibitor 2A (melanoma p16 inhibits CDK4) antibody
    • Cyclin Dependent Kinase Inhibitor 2A antibody
    • Cyclin dependent kinase inhibitor 2A isoform 4 antibody
    • Cyclin dependent kinase inhibitor 2A isoforms 1/2/3 antibody
    • Cyclin dependent kinase inhibitor p16 antibody
    • INK4 antibody
    • INK4A antibody
    • MLM antibody
    • MTS1 antibody
    • Multiple tumor suppressor 1 antibody
    • p14 antibody
    • p16 antibody
    • P16INK4 antibody
    • p16INK4a antibody
    • p19 antibody
    • p19Arf antibody
    • TP16 antibody
    see all

Images

  • All lanes : Anti-CDKN2A/p16INK4a antibody [EPR1473] (ab108349) at 1/2000 dilution (purified)

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
    Lane 2 : Saos-2 (Human osteosarcoma cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 17 kDa
    Observed band size: 17 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 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.

  • 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.

  • 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.

  • 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.

     

  • Anti-CDKN2A/p16INK4a antibody [EPR1473] (ab108349) at 1/2000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 17 kDa
    Observed band size: 17 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-CDKN2A/p16INK4a antibody [EPR1473] (ab108349) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney) cell lysate
    Lane 3 : Saos-2 (Human osteosarcoma cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 17 kDa

  • 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.

  • 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.

References

This product has been referenced in:
  • Li X  et al. Upregulation of lactate-inducible snail protein suppresses oncogene-mediated senescence through p16INK4ainactivation. J Exp Clin Cancer Res 37:39 (2018). Read more (PubMed: 29482580) »
  • Wang H  et al. Mangiferin induces islet regeneration in aged mice through regulating p16INK4a. Int J Mol Med 41:3231-3242 (2018). Read more (PubMed: 29512742) »

See all 36 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (growth plate)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: hyaluronidase
Permeabilization
No
Specification
growth plate
Blocking step
Bloxall then 10% Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 23°C
Fixative
zinc buffered formallin
Username

Abcam user community

Verified customer

Submitted Mar 12 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (vascular smooth muscle celss)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
30 µg
Specification
vascular smooth muscle celss
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 21 2017

Application
Western blot
Sample
Human Cell lysate - nuclear (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jul 04 2017

Application
ELISA
Sample
Human Recombinant protein (Purified recombinant p16 (overexpressed in HEK293))
Specification
Purified recombinant p16 (overexpressed in HEK293)
Type
Sandwich (Detection)
Blocking step
SuperBlock PBS Pierce #37580 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10µg/mL · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Apr 28 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (B16F10 melanoma cancer implanted)
Specification
B16F10 melanoma cancer implanted
Fixative
4% Formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: autoclaved for 10min at 121 Celcius (10mM Sodium Citrate, pH 6.0)
Permeabilization
Yes - 0.1% SDS
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Ms. Seontae Kim

Verified customer

Submitted Mar 15 2013

Thank you for contacting us.

We do not provide Abtrial code for tested species and applications. Please check and explain this to customer;

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborati...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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