Human p16INK4a / CDKN2A Antibody

(3 citations)
(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human p16INK4a/CDKN2A in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human p16INK4a/CDKN2A
    Glu2-Asp156
    Accession # P42771
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunocytochemistry
    0.3-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human p16INK4a/CDKN2A by Western Blot. 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.
Immunocytochemistry
p16INK4a / CDKN2A in HeLa Human Cell Line. 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.
Detection of Human p16INK4a / CDKN2A by Simple WesternTM. 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 the
12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: p16INK4a / CDKN2A
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.
  • Long Name:
    p16 Cyclin Dependent Kinase 4 Inhibitor 2A
  • Entrez Gene IDs:
    1029 (Human); 12578 (Mouse); 25163 (Rat)
  • Alternate Names:
    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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. MAX inactivation is an early event in GIST development that regulates p16 and cell proliferation
    Authors: 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
    Nat Commun, 2017;8(0):14674.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Adiponectin corrects premature cellular senescence and normalizes antimicrobial peptide levels in senescent keratinocytes
    Authors: Taewon Jin
    Biochem Biophys Res Commun, 2016;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Induction of heparanase by HPV E6 oncogene in head and neck squamous cell carcinoma.
    Authors: Hirshoren N, Bulvik R, Neuman T, Rubinstein A, Meirovitz A, Elkin M
    J Cell Mol Med, 2014;18(1):181-6.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
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Normal Goat IgG Control

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Goat IgG HRP-conjugated Antibody

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Goat IgG HRP-conjugated Antibody

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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

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Donkey Anti-Goat IgG Antibody

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Donkey Anti-Goat IgG Biotinylated Antibody

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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Western Blot Human p16INK4a / CDKN2A Antibody AF5779
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Very Good
 WB Human Anonymous 05/20/2016
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Western Blot Human p16INK4a / CDKN2A Antibody AF5779
Western Blot: Human p16INK4a / CDKN2A Antibody [AF5779]

Summary

ApplicationWestern Blot
Sample TestedMesenchymal stem cells
SpeciesHuman

Other Experimental Details

Other Experimental Details5% non fat milk as blocking agent for 1 hour and overnight incubation with primary antibody at 1:3000. Specific band at 16kDa as expected, but also band at 25 KDa. Cells at passage 2 had low p16 expression, while at passage 6 expression was increased as expected. Loading: p2,p6-p2,p6-p2,p6-p2,p6 of MSCs from 4 different individuals.

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