Anti-CDKN2A/p16INK4a antibody [DCS50.1] (ab16123)

Overview

  • Product nameAnti-CDKN2A/p16INK4a antibody [DCS50.1]See all CDKN2A/p16INK4a primary antibodies ...
  • Description
    Mouse monoclonal [DCS50.1] to CDKN2A/p16INK4a
  • SpecificityThis antibody shws no cross reactivity with the closely related inhibitors p15INK4b and p18INK4c.
  • Tested applicationsICC/IF, Sandwich ELISA, WB, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human CDKN2A/p16INK4a aa 1-156.
    Sequence:

    MEPAAGSSMEPSADWLATAAARGRVEEVRALLEAGALPNAPNSYGRRPIQ VMMMGSARVAELLLLHGAEPNCADPATLTRPVHDAAREGFLDTLVVLHRA GARLDVRDAWGRLPVDLAEELGHRDVARYLRAAAGGTRGSNHARIDAAEG PSDIPD


    Database link: P42771

  • Positive control
    • HEK293 cells, HeLa ((Human epithelial carcinoma cell line).

Properties

Applications

Our Abpromise guarantee covers the use of ab16123 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
ICC/IF Use a concentration of 1 µg/ml.
Sandwich ELISA Use a concentration of 1 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to CDKN2A/p16INK4a (ab7962). For sandwich ELISA, use this antibody as Capture at 1 µg/ml with Rabbit polyclonal to CDKN2A/p16INK4a (ab7962) as Detection.
WB Use at an assay dependent dilution. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IP Use at an assay dependent dilution. Ab16123 co-precipitates cdk4/cdk6.
Flow Cyt Use 1µg for 106 cells.

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 [DCS50.1] images

  • ICC/IF image of ab16123 stained Hepp cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16123, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) ab150113) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HEK293 cells stained with ab16123 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16123, 1µg/1x106 cells ) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 100% methanol used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

  • All lanes : Anti-CDKN2A/p16INK4a antibody [DCS50.1] (ab16123) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 16 kDa
    Observed band size : 16 kDa
    Additional bands at : 37 kDa,50 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded bladded tumor tissue labelling CDKN2A/p16INK4a with ab16123 at 10µg/ml.

  • Standard Curve for CDKN2A/p16INK4a (Analyte: CDKN2A/p16INK4a protein (Human) (ab84075)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [DCS50.1] to CDKN2A/p16INK4a (ab16123) at 1µg/ml and Detector Antibody Rabbit polyclonal to CDKN2A/p16INK4a (ab7962) at 0.5µg/ml.

Protocols

References for Anti-CDKN2A/p16INK4a antibody [DCS50.1] (ab16123)

ab16123 has not yet been referenced specifically in any publications.

Product Wall

Application ELISA
Sample Human Recombinant protein (Purified recombinant p16 (overexpressed in HEK293))
Specification Purified recombinant p16 (overexpressed in HEK293)
Type Sandwich (Capture)
Blocking step SuperBlock PBS Pierce #37580 as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10µg/mL
Username

Abcam user community

Verified customer

Submitted Apr 28 2014

Thank you for contacting us.

Abcam takes product quality seriously and we recently undertook an in-depth study examining storage conditions and antibody performance. After over 13,000 individual ELISA experiments, we have determined that sto...

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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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Thank you for contacting us.

Unfortunatley we do not calculate the isoelectric point of our research antibodies.

I hope this information is neverthelesshelpful to you. Please do not hesitate to contact us if you need any more advi...

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I am very pleased to hear you would like to accept our offer and test ab16123 in mouse in Flow cytometry. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview formouse in Flow cytom...

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Thank you for your email. I am very sorry to hear that the replacement antibody Carolyn sent you also did not work. I certainly understand that your time is valuable and I would be happy to send you another anti-p16 antibody. I checked our current stoc...

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Thank you for taking time to complete our questionnaire. I am sorry to hear that antibodies ab16123, ab39184 and ab61224 are not providing satisfactory results. The details provided will enable us to investigate these cases and will provide us with ...

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Thank you for contacting us. We have indeed in our catalog 5 unconjugated anti-interferon alpha 1 antibodies tested and guaranteed to work in Western Blot and Human samples. Please note that, unfortunately, import restrictions make the ...

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Application Western blot
Sample Cow Cell lysate - whole cell (vascular endothelial cells)
Loading amount 20 µg
Specification vascular endothelial cells
Gel Running Conditions Reduced Denaturing (15%)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 22 2011

Thank you for your enquiry. I am sorry it has taken a little time to obtain this information for you. Thank you for your patience. I can confirm that p16 is widely expressed. This antibody has been used successfully in western blotting with HeL...

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