Anti-CDKN2A/p16INK4a antibody [2D9A12] (ab54210)

Overview

  • Product nameAnti-CDKN2A/p16INK4a antibody [2D9A12]See all CDKN2A/p16INK4a primary antibodies ...
  • Description
    Mouse monoclonal [2D9A12] to CDKN2A/p16INK4a
  • Tested applicationsFlow Cyt, WB, ELISA, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human CDKN2A/p16INK4a .
    Database link: Q8N726

  • Positive control
    • Rat liver tissue; Human brain tissue; Human brain tumor tissue; PFA/Tween HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab54210 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
Flow Cyt 1/50.
WB 1/500 - 1/2000. Tested using a recombinant protein expressed in E. coli. We have no data on the detection of the endogenous protein and would welcome feedback from anyone that has performed a WB with ab54210 using cell lysates.
ELISA 1/10000.
ICC/IF Use at an assay dependent concentration. PubMed: 24828678
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Recommended antigen retrieval: Place slides in a special glass slide holder and fill in the rack with slides to ensure even heating. Rinse rack in 1L of Citrate buffer in ordinary pressure cooker,(0.01M,pH6.0) to immerse all slides. Cover the cap and heat for 15-20 minutes on electric cooker until water boils. Begin timing, only 2 minutes in high-pressure Remove the pressure cooker, Allow it to cool(Cool the slides at room temperature) Rinse the slides in PBS for 5 minutes

Target

  • Cellular localizationCytoplasmic and Nuclear
  • Database links
  • FormThere 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
    • ARF antibody
    • CCM2 antibody
    • CDK4 Inhibitor 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
    • INK4 antibody
    • INK4A antibody
    • MLM antibody
    • MTS1 antibody
    • Multiple tumor suppressor 1 antibody
    • Multiple Tumor Supressor 1 antibody
    • p14 antibody
    • p14(ARF) antibody
    • p14ARF antibody
    • p16 antibody
    • p16 INK4 antibody
    • p16INK4 antibody
    • P16INK4 antibody
    • p16INK4a antibody
    • p19 antibody
    • p19(ARF) antibody
    • p19Arf antibody
    • TP16 antibody
    see all

Anti-CDKN2A/p16INK4a antibody [2D9A12] images

  • Overlay histogram showing HeLa cells stained with ab54210 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. 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 (ab54210, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • All lanes : Anti-CDKN2A/p16INK4a antibody [2D9A12] (ab54210) at 1/500 dilution

    Lane 1 : Molecular weight marker
    Lane 2 : Truncated p16 recombinant protein


    Observed band size : 40 kDa (why is the actual band size different from the predicted?)
  • ab54210 (1:500) staining p16 INK4a in rat liver tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.

  •  ab54210 (1:500) staining p16 INK4a in human brain tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.

  •  ab54210 (1:500) staining p16 INK4a in human brain tumor tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.

References for Anti-CDKN2A/p16INK4a antibody [2D9A12] (ab54210)

This product has been referenced in:
  • Qin S  et al. Homeostatic maintenance of allele-specific p16 methylation in cancer cells accompanied by dynamic focal methylation and hydroxymethylation. PLoS One 9:e97785 (2014). ICC/IF ; Human . Read more (PubMed: 24828678) »
  • Zhang B  et al. SIRT3 overexpression antagonizes high glucose accelerated cellular senescence in human diploid fibroblasts via the SIRT3-FOXO1 signaling pathway. Age (Dordr) 35:2237-53 (2013). WB ; Human . Read more (PubMed: 23494737) »

See all 9 Publications for this product

Product Wall



CDKN2A/p16INK4a est une protéine qui est exprimé dans 63% de tumeurs et non pas que dans les tumeurs cérébrales. Vous pouvez donc également utiliser d’autres tissus cancéreux comme contrôl...

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Thank you for contacting us.

Human brain slides, such as ab4304, or humor brain tumor slides such as ab4679 should be good positive controls.

I hope this information is helpful to you. Please do not hesitate to contact us if you need ...

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Thank you for contacting us.

Here are the links to the Human Protein Atlas, including ones to the sites for normal and cancer tissue as well as cell lines. Several brain cell lines are listed as expressing CDKN2A in moderate levels.

h...

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Thank you for contacting Abcam and it was good to meet you at the Distributor Day events.

I have looked through our catalogue to try and find some antibodies that would match your customers needs and I have placed links to the best matches bel...

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Thank you for your enquiry and your interest in our products.
We have several antibodies against CDKN2A but none of them have been tested for cross-reactivity with Syrian hamster (Mesocricetus auratus).
Regarding ab54210 - the exact immunogen...

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Thank you for taking time tocontact us.

For ab16123, have you tried running a positive control with HeLa cell lysate? It may be possible that your cell line is expressing a different isoform. In our testing, we do see the predominant band at ...

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Application Western blot
Sample Rat Tissue lysate - whole (heart (left ventricle))
Loading amount 20 µg
Specification heart (left ventricle)
Gel Running Conditions Reduced Denaturing (15% poly-acrylamide)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Jun 07 2012

Thank you for your patience in awaiting our reply, the person who did the testing was away for several days. The testing was done at a 1/50 dilution. The datasheet is being updated to reflect this. Please let me know if you have any further questions.

Thank you for contacting us. The concentration for this product is not determined as it is from ascites. Concentration values are only meaning for purified antibodies, since those from ascites may contain proteins other than the protein of interest. Th...

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Thank you for your phone call. Please find the reference I mentioned attached. I hope this helps, please let me know if you need any other information or assistance.

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