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

概述

  • 产品名称
    Anti-CDKN2A/p16INK4a抗体[2D9A12]
    参阅全部 CDKN2A/p16INK4a 一抗
  • 描述
    小鼠单克隆抗体[2D9A12] to CDKN2A/p16INK4a
  • 宿主
    Mouse
  • 经测试应用
    适用于: Flow Cyt, ELISA, IHC-Pmore details
    不适用于: WB
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Recombinant fragment corresponding to Human CDKN2A/p16INK4a.
    Database link: P42771

  • 阳性对照
    • IHC-P: Human brain tumor, ovarian carcinoma, cervix, skin and brain tissue. Rat liver tissue. Flow Cyt: HeLa cells.
  • 常规说明

    We have received positive and negative feedback from customers for ICC/IF. ICC/IF is not a batch-tested application for this antibody and we are not able to cover it under our Abpromise guarantee.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt 1/50.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ELISA 1/10000.
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
  • 应用说明
    Is unsuitable for WB.
  • 靶标

    • 细胞定位
      Cytoplasmic and Nuclear
    • 数据库链接
    • 形式
      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.
    • 别名
      • 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

    图片

    • Characterization of methylation and expression of p16 in the fusion cell lines.

      Confocal immunocytochemistry staining shows varying P16 expression in the various cell lines.

      Fusion cells were fixed in 4% formaldehyde/PBS for 10 minutes before a 5-minute wash in PBS. Fixed samples were blocked for 30 minutes with 0.5% Triton X-100/PBS at room temperature. Cells were incubated with ab54210 for 1 hour at 37°C, after washed with 0.5% Triton X-100/PBS, then incubated with goat-anti rabbit IgG antibody labeled with rhodamine for 1 hour at 37°C DAPI (1∶2000) was used to stain the cell nucleus.

      Photos were taken on a Leica confocal microscope.

    • ab54210 staining CDKN2A/p16INK4a in human cervix and ovarian carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

      Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/4000) for 20 minutes at 25°C. An undiluted HRP-conjugated mouse polymer was used as the secondary antibody.

      See Abreview

    • ab54210 staining CDKN2A/p16INK4a in human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

      Tissue was fixed with formaldehyde and blocked with 2% BSA for 1 hour at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in 1% NGS/TBS) for 20 hours at 4°C. ab6785 (1/800) was used as the secondary antibody.

      See Abreview

    • ab54210 at a 1:500 staining CDKN2A/p16INK4a in human brain tumor tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (DAB) is shown.

    • ab54210 at a 1:500 staining CDKN2A/p16INK4a in rat liver tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (DAB staining) is shown.

    • ab54210 at a 1:500 staining CDKN2A/p16INK4a in human brain tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (DAB staining) is shown.

    • Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) 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.

    文献

    This product has been referenced in:
    • Kehm R  et al. Age-related oxidative changes in pancreatic islets are predominantly located in the vascular system. Redox Biol 15:387-393 (2018). Read more (PubMed: 29331666) »
    • Tsuboi T  et al. Administration of L-arginine plus L-citrulline or L-citrulline alone successfully retarded endothelial senescence. PLoS One 13:e0192252 (2018). Read more (PubMed: 29415069) »

    See all 29 Publications for this product

    客户评价及客户问答

    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 ...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (heart (left ventricle))
    Gel Running Conditions
    Reduced Denaturing (15% poly-acrylamide)
    Loading amount
    20 µg
    Specification
    heart (left ventricle)
    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

    提交于 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.

    Thank you for contacting Abcam.
    Your credit note ID is *********.
    I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask yo...

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    Thank you for contacting Abcam.

    Unfortunately for this antibody, the expression vector was provided for us for protein expression from another sourceto be used as the antigen. We were never supplied with the protein sequence for it as that in...

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    Thank you for your telephone enquiry yesterday afternoon.

    As discussed on the phone, I am sorry to confirm that to our knowledge, ab54210 has not been testedin ICC-IF. All testedand guaranteedapplications and species are listed on the online...

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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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