Human p16INK4a / CDKN2A Antibody

R&D Systems | Catalog # AF5779

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all p16INK4a / CDKN2A Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human p16INK4a/CDKN2A
Glu2-Asp156
Accession # P42771

Specificity

Detects human p16INK4a/CDKN2A in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human p16INK4a / CDKN2A Antibody

Detection of Human p16INK4a/CDKN2A antibody by Western Blot.

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.
p16INK4a / CDKN2A antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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 antibody by Simple WesternTM.

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.
Detection of p16INK4a/ CDKN2A by Western Blot

Detection of p16INK4a/ CDKN2A by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/35563794), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of p16INK4a/ CDKN2A by Western Blot

Detection of p16INK4a/ CDKN2A by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/35563794), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human p16INK4a / CDKN2A Antibody

Application
Recommended Usage

Immunocytochemistry

0.3-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Simple Western

10 µg/mL
Sample: HEK293 human embryonic kidney cell line

Western Blot

1 µg/mL
Sample: HEK293 human embryonic kidney cell line, HepG2 human hepatocellular carcinoma cell line, and Saos-2 human osteosarcoma cell line

Reviewed Applications

Read 2 reviews rated 4.5 using AF5779 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

ARF, CDK4I, CDKN2A, INK4a, MLM, p14ARF, p16, p19ARF

Entrez Gene IDs

1029 (Human); 12578 (Mouse); 25163 (Rat)

Gene Symbol

CDKN2A

UniProt

P42771

Additional p16INK4a / CDKN2A Products

Product Documents for Human p16INK4a / CDKN2A Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human p16INK4a / CDKN2A Antibody

For research use only

Citations for Human p16INK4a / CDKN2A Antibody

Customer Reviews for Human p16INK4a / CDKN2A Antibody (2)

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  • Human p16INK4a / CDKN2A Antibody
    Name: Nicole Malet
    Application: Simple Western
    Sample Tested: HEK293 human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 08/12/2021
    Human p16INK4a/CDKN2A Antibody Ref:AF5779 R et D Systems Efficacité validée dans conditions suivantes: Dilution anticorps p16= 1/10è (0,2mg/ml solution stock) [HEK]= 0,65mg/ml Anticorps 2aire goat prêt à l’emploi
    Human p16INK4a / CDKN2A Antibody AF5779
  • Human p16INK4a / CDKN2A Antibody
    Name: Juliana Redondo
    Application: Western Blot
    Sample Tested: Mesenchymal stem cells
    Species: Human
    Verified Customer | Posted 05/20/2016
    5% 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.
    Human p16INK4a / CDKN2A Antibody AF5779

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