p16INK4a/CDKN2A Antibody [Unconjugated]

Western blot shows lysates of HEK293 human embryonic kidney cell line, HepG2 human hepatocellular carcinoma cell line, and Saos-2 human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human p16INK4a/CDKN2A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5779) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for p16INK4a/CDKN2A at approximately 16 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

p16INK4a / CDKN2A was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human p16INK4a / CDKN2A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5779) at 0.3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of HEK293 human embryonic kidney cell line, loaded at 0.2 mg/mL. A specific band was detected for p16INK4a / CDKN2A at approximately 24 kDa (as indicated) using 10 µg/mL of Goat Anti-Human p16INK4a / CDKN2A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5779) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.

Protein markers of cellular senescence. Western blot analysis of (A) NHEM transduced with mutated BRAFV600E (all n = 7, p16 n = 6) and (B) melanoma cell line Mel Juso treated with 100 µM etoposide (n = 3). (C,D) Immunofluorescent stainings of PML and DAPI. Scale bars equal 20 µm. Bars are shown as mean ± SEM (Student’s t-test). (*: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35563794), licensed under a CC-BY license. Not internally tested by R&D Systems.

Protein markers of cellular senescence. Western blot analysis of (A) NHEM transduced with mutated BRAFV600E (all n = 7, p16 n = 6) and (B) melanoma cell line Mel Juso treated with 100 µM etoposide (n = 3). (C,D) Immunofluorescent stainings of PML and DAPI. Scale bars equal 20 µm. Bars are shown as mean ± SEM (Student’s t-test). (*: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35563794), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

p16INK4a/CDKN2A Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human p16INK4a/CDKN2A
  Glu2-Asp156
  Accession # P42771
• Specificity: Detects human p16INK4a/CDKN2A in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CDKN2A
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 0.3-15 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for p16INK4a/CDKN2A Antibody [Unconjugated]

• ARF
• CDK4 inhibitor p16-INK4
• CDK4I
• CDK4IP14ARF
• CDKN2
• CDKN2A
• cell cycle negative regulator beta
• CMM2P16-INK4A
• Cyclin-dependent kinase 4 inhibitor A
• cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4)
• cyclin-dependent kinase inhibitor 2A
• INK4
• INK4a
• MLM
• MLMP16INK4
• MTS-1
• MTS1P14
• Multiple tumor suppressor 1
• p14
• p14ARF
• p16
• p16-INK4
• p16INK4a
• p16-INK4a
• P19
• p19ARF
• TP16

Background

p16INK4a (16 kDa Inhibitor of CDK4-a; also MTS1, CDK41 and CDKN2) is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or 6, thereby blocking cyclin binding and subsequent Ser/Thr kinase activity. Human p16INK4a is 156 amino acids (aa) in length. It contains four “L” shaped ankyrin repeats
(aa 11‑139) that interact with cyclin. There are at least two splice variants for p16INK4a. One is termed p12 and shows a 65 aa substitution for aa 52‑156; the other simply shows an alternate start site at Met52. Full‑length human p16INK4a shares 63% aa identity with mouse p16INK4a.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for p16INK4a/CDKN2A Antibody (AF5779)(14)

Publications using AF5779

• Featherby, SJ;Faulkner, EC;Ettelaie, C; Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation Molecular medicine reports 2025-02-01 [PMID: 39611476] (Western Blot, Human)
• JR Herdy, L Traxler, RK Agarwal, L Karbacher, JCM Schlachetz, L Boehnke, D Zangwill, D Galasko, CK Glass, J Mertens, FH Gage Increased post-mitotic senescence in aged human neurons is a pathological feature of Alzheimer&#039;s disease Cell Stem Cell, 2022-12-01;29(12):1637-1652.e6. 2022-12-01 [PMID: 36459967] (Simple Western, Human)
• Tom Zimmermann, Michaela Pommer, Viola Kluge, Chafia Chiheb, Susanne Muehlich, Anja-Katrin Bosserhoff Detection of Cellular Senescence in Human Primary Melanocytes and Malignant Melanoma Cells In Vitro Cells 4/28/2022 [PMID: 35563794]
• V Farfariell, DV Gordienko, L Mesilmany, Y Touil, E Germain, I Fliniaux, E Desruelles, D Gkika, M Roudbaraki, G Shapovalov, L Noyer, M Lebas, L Allart, N Zienthal-G, O Iamshanova, F Bonardi, M Figeac, W Laine, J Kluza, P Marchetti, B Quesnel, D Metzger, D Bernard, JB Parys, L Lemonnier, N Prevarskay TRPC3 shapes the ER-mitochondria Ca2+ transfer characterizing tumour-promoting senescence Nature Communications, 2022-02-17;13(1):956. 2022-02-17 [PMID: 35177596] (Western Blot, Human)
• SL Mathiasen, L Gall-Mas, IS Pateras, SDP Theodorou, MRJ Namini, MB Hansen, OCB Martin, CK Vadivel, K Ntostoglou, D Butter, M Givskov, C Geisler, AN Akbar, VG Gorgoulis, T Frisan, N Ødum, T Krejsgaard Bacterial genotoxins induce T�cell senescence Cell Reports, 2021-06-08;35(10):109220. 2021-06-08 [PMID: 34107253] (Western Blot, Human)
• Rachael E. Schwartz, Maxim N. Shokhirev, Leonardo R. Andrade, J. Silvio Gutkind, Ramiro Iglesias-Bartolome, Gerald S. Shadel Insights into epithelial cell senescence from transcriptome and secretome analysis of human oral keratinocytes Aging (Albany NY) 2/28/2021 [PMID: 33601339]
• P Sachamitr, JC Ho, FE Ciamponi, W Ba-Alawi, FJ Coutinho, P Guilhamon, MM Kushida, FMG Cavalli, L Lee, N Rastegar, V Vu, M Sánchez-Os, J Coulombe-H, E Kanshin, H Whetstone, M Durand, P Thibault, K Hart, M Mangos, J Veyhl, W Chen, N Tran, BC Duong, AM Aman, X Che, X Lan, O Whitley, O Zaslaver, D Barsyte-Lo, LM Richards, I Restall, A Caudy, HL Röst, ZQ Bonday, M Bernstein, S Das, MD Cusimano, J Spears, GD Bader, TJ Pugh, M Tyers, M Lupien, B Haibe-Kain, H Artee Luch, S Weiss, KB Massirer, P Prinos, CH Arrowsmith, PB Dirks PRMT5 inhibition disrupts splicing and stemness in glioblastoma Nature Communications, 2021-02-12;12(1):979. 2021-02-12 [PMID: 33579912] (Western Blot, Human)
• Ruizhu Lin, Lea Rahtu-Korpela, Johanna Magga, Johanna Ulvila, Julia Swan, Anna Kemppi et al. miR-1468-3p Promotes Aging-Related Cardiac Fibrosis Molecular Therapy - Nucleic Acids 6/5/2020 [PMID: 32348937]
• P Vijayaraj, A Minasyan, A Durra, S Karumbayar, M Mehrabi, CJ Aros, SD Ahadome, DW Shia, K Chung, JM Sandlin, KF Darmawan, KV Bhatt, CC Manze, MK Paul, DC Wilkinson, W Yan, AT Clark, TM Rickabaugh, WD Wallace, TG Graeber, R Damoiseaux, BN Gomperts Modeling Progressive Fibrosis with Pluripotent Stem Cells Identifies an Anti-fibrotic Small Molecule Cell Rep, 2019-12-10;29(11):3488-3505.e9. 2019-12-10 [PMID: 31825831] (ICC, Western Blot, Human)
• Jianjun Wang, Yongliang Yao, Qinghui Zhang, Shasha Li, Lijun Tang Inflammatory responses induced by Helicobacter pylori on the carcinogenesis of gastric epithelial GES‑1 cells International Journal of Oncology 4/9/2019 [PMID: 31081048]
• SY Choi, BH Bin, W Kim, E Lee, TR Lee, EG Cho Exposure of human melanocytes to UVB twice and subsequent incubation leads to cellular senescence and senescence-associated pigmentation through the prolonged p53 expression J. Dermatol. Sci., 2018-03-02;0(0):. 2018-03-02 [PMID: 29525471] (Western Blot, Human)
• IM Schaefer, Y Wang, CW Liang, N Bahri, A Quattrone, L Doyle, A Mariño-Enr, A Lauria, M Zhu, M Debiec-Ryc, S Grunewald, JF Hechtman, A Dufresne, CR Antonescu, C Beadling, ET Sicinska, M van de Rij, GD Demetri, M Ladanyi, CL Corless, MC Heinrich, CP Raut, S Bauer, JA Fletcher MAX inactivation is an early event in GIST development that regulates p16 and cell proliferation Nat Commun, 2017-03-08;8(0):14674. 2017-03-08 [PMID: 28270683] (Western Blot, Human)
• Taewon Jin Adiponectin corrects premature cellular senescence and normalizes antimicrobial peptide levels in senescent keratinocytes Biochem Biophys Res Commun, 2016-06-25;0(0):. 2016-06-25 [PMID: 27349869] (Western Blot, Human)
• Hirshoren N, Bulvik R, Neuman T, Rubinstein A, Meirovitz A, Elkin M Induction of heparanase by HPV E6 oncogene in head and neck squamous cell carcinoma. J Cell Mol Med, 2013-11-28;18(1):181-6. 2013-11-28 [PMID: 24286246] (IHC, Human)

Bioinformatics

• Gene Symbol: CDKN2A
• Uniprot:
  • P42771()