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Anti-CDKN2A Antibody EP4353Y(3)
Also for CDKN2A (NM_000077)
|A full length p16/INK4A protein with GST tag was used as an immunogen.|
||Tissue culture supernatant
||Lot dependent; please refer to CoA along with shipment
|WB, IF, FC
||WB: 1:2000; ICC/IF: 1:100 - 1:500; FC: 1:50 - 1:100
|Does not react with Mouse, Rat. Is unsuitable for IHC-P or IP.|
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
|Is unsuitable for IHC-P or IP.
|Homo sapiens cyclin-dependent kinase inhibitor 2A (CDKN2A), transcript variant 1|
|ARF; CDK4I; CDKN2; CMM2; INK4; INK4A; MLM; MTS-1; MTS1; P14; P14ARF; P16; P16-INK4A; P16INK4;|
|The division cycle of eukaryotic cells is regulated by a family of protein kinases known as the cyclin-dependent kinases (CDKs). The sequential activation of individual members of this family and their consequent phosphorylation of critical substrates promotes orderly progression through the cell cycle. It has been reported that p16 binds to CDK4 and inhibits the catalytic activity of the CDK4/cyclin D enzymes. p16 seems to act in a regulatory feedback circuit with CDK4, D-type cyclins and retinoblastoma protein (1). The INK4 (inhibitor of cyclin-dependent kinase 4) family consists of four tumor-suppressor proteins: p15(INK4B), p16(INK4A), p18(INK4C), and p19(INK4D). While their sequences and structures are highly homologous, they show appreciable differences in conformational flexibility, stability, and aggregation tendency (2). Cell cycle arrest at the G1 checkpoint allows completion of critical macromolecular events prior to S phase. Regulators of the G1 checkpoint include an inhibitor of cyclin-dependent kinase, p16INK4; two tumor-suppressor proteins, p53 and RB and cyclin D1. p16INK4 is a tumor-suppressor protein and that genetic and epigenetic abnormalities in genes controlling the G1 checkpoint can lead to both escape from senescence and cancer formation (3). |
Senescence and Autophagy
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Western blot - CDKN2A/p16INK4a antibody [EP4353Y]; All lanes : Anti-CDKN2A/p16INK4a antibody [EP4353Y(3)] at 1/20000 dilution.Lane 1 : HeLa cell lysate.Lane 2 : 293 cell lysate.Lysates/proteins at 10 µg per lane.Secondary.HRP labelled goat anti rabbit at 1/2000 dilution.Predicted band size : 17 kDa.Observed band size : 17 kDa.
Immunocytochemistry/ Immunofluorescence - CDKN2A/p16INK4a antibody [EP4353Y]; Immunofluorescent staining of HeLa cells using 1/100 TA300862
Flow Cytometry-Anti-CDKN2A/p16INK4a antibody [EP4353Y](TA300862); Overlay histogram showing HEK293 cells stained with TA300862 (red line). The cells were fixed with 4% paraformaldehyde (10 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 for 30 min at 22Â°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22Â°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1Âµg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.