Description
Cyclins and cyclin-dependent kinases (cdks) form active complexes that regulate key events during the progression of the cell cycle and are evolutionarily highly conserved. The p16 protein has been identified as a specific inhibitor of cdk4 because it blocks cdk4 substrate phosphorylation. p16 inhibits cdk4 dependent phosphorylation of the tumor suppressor retinoblastoma protein (Rb) and Rb related proteins, p107 and p130. The biochemical properties of p16 suggest that it may be a tumor suppressor gene product. Recently a gene cloned from the short arm of human chromosome 9, Multiple Tumor Suppressor 1 (MTS1) has been identified as the gene for p16. The gene, now also known as the CDKN2 gene, has been found to be mutated in a very high percentage of tumors, including 75% of melanoma cell lines.
Western blot: For protocols on Western blotting, please refer to "Cell Biology (WB, IP, IHC, IF)" at our website: http://www.bdbiosciences.com/us/s/resources Endogenous p16 has been detected in several cell lines including Saos-2 human osteosarcoma (ATCC HTB-85), WI-38 human lung fibroblast (ATCC CCL-75), HeLa cervical carcinoma (ATCC CCL-2) and 293 adenovirus immortalized human kidney (ATCC CRL-1573) cells. U-2 OS human osteosarcoma (ATCC HTB-96) and MCF7 human breast carcinoma (ATCC HTB-22) have been reported to have undetectable levels of p16 and are suggested as negative controls.
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. An isotype control should be used at the same concentration as the antibody of interest.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
