Detection of p53 in Camptothecin-treated MCF‑7 Human Cell Line by Flow Cytometry.
MCF‑7 human breast cancer cell line was treated with Camptothecin overnight (filled histogram) or untreated (open histogram), then stained with Mouse Anti-Human Phospho-p53 (S15) PE‑conjugated Monoclonal Antibody (Catalog # IC1839P) or isotype control antibody (Catalog # IC002P, data not shown). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
p53 is a 52-55 kDa member of the p53 family of
proteins. It is ubiquitously expressed,
being found in both cytoplasm (as a monomer) and nucleus (as an
oligomer/tetramer). Human p53 is 393
amino acids in length and contains multiple domains that interact with multiple
binding partners. There are numerous
posttranslational modification sites, including multiple phosphorylation,
acetylation, NEDDylation and ubiquitination sites, plus one SUMOylation and two
methylation sites. N-terminal
modifications tend to stabilize p53, while C-terminal modifications impact
molecule oligomerization, DNA-binding and ubiquitination. Phosphorylation on Ser15 inhibits MAM2
modification of p53, thereby extending its half-life. Notably, there are nine possible p53 isoforms,
six of which do not contain Ser15 of the standard (or full-length) form. Thus, they are unlikely to show strong positive
staining that is associated with the phosphorylation of Ser15. p53 is best known as a preserver of cell
genome integrity. In the nucleus, it stimulates needed repairs to DNA and, if
necessary, cease cell division and/or initiate apoptosis. In addition, it has at least one role in the
cytoplasm where it is known to regulate glucose consumption.
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