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 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
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.
Western Blot: Human p16INK4a / CDKN2A Antibody [AF5779].
Mesenchymal stem cells
Other Experimental Details
Other Experimental Details
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.