Anti-p53 p-Ser315 (human) antibody, mouse monoclonal (#18)
Product code71-117Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium.

ImmunogenSynthetic peptide containing phosphor-Ser315 of p53

Isotype

Mouse IgG2bκ

Reactivity

HumanSpecial notes

N/A

Application

1. Western blotting (x 1,000~2000 dilution, Fig.1)
2. Immunohistochemistry (Fig.2)
3. Immunofluorescence staining (Fig.3)
4. ELISA

Background

p53 mutants are found in more than half of human cancers and are consideredas the most important human cancer related gene. p53 is detected at 53kD
position by electrophoresis and is composed of 393 amino acids.

In theunstressed normal cells, the p53 level is low and it is inactive. However, withstress, especially with DNA damage, it is activated to promote arrest of cell cycle
and repair of DNA damage, or induction of apoptosis.

The functions andstability of p53 are regulated by phosphorylation of serine and threonine, andacetylation of lysine at various sites in the molecule.
Ser315 is phosphorylated by aurora kinase and cycline-dependent kinases whencells are subjected to stress such as DNA damage and microtubule disruption bynocodazole (ref 1, 2 & 3).

However the effect of the phosphorylation on thefunction of p53 is mostly unknown.
This product is the purified IgG fraction obtained from serum-free culture
medium of mouse hybridoma (clone #18) which produces monoclonal antibody
that specifically recognizes human p53 protein with phospholyrated Ser315.
Data Link

UniProtKB/Swiss-Prot P04637 (P53_HUMAN)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 71-117 Anti-p53 p-Ser315 (human) antibody, mouse monoclonal (#18)

(kD)
220

1

2120
100
80
60

p53(ser31p53(p-S315)

50

55

40

IB:anti-p53(p-S315)(kD)
80

ser315

60p53

50
40

IB:anti-p53

Western blotting.
Sample: Crude cell extracts of MCF7 untreated (lane 1) and treated with nocodazole at 100 ng/ml for48 h (lane2).
The lower panel is the whole p53 protein identified by omnipotent anti-p53 antibody (DO-1).

Fig.2 Immunohistochemistry of stomach cancer.
(Formalin/PFA-fixed paraffin-embedded section)

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Fig.3. Immunofluorescence staining of p53 phosho-Ser315 in nuclei of HeLa cells subjected DNA
damage.
1. HeLa cells were treated with 100 nM Doxorubicin for 24 hr, fixed with 4% paraformaldehyde
overnight, permealized with 0.25% Triton X-100 in PBS for 10 min.
2.

Incubate cells with 1.5% BSA in PBS for 30 min to block unspecific binding of the antibodies.
Incubate the cells with 1/1,000 diluted anti-p53(p-315) antibody in 1% BSA in PBS at 4 ℃
overnight.

3. Incubate cells with the secondary antibody, goat anti-mouse IgG conjugated with Alex 488, at
1/1,000 dilution in 1% BSA for 1 hr at room temperature.
4. Nucleus (DNA) was stained with DAPI
References
1. Katayama H et al “Phosphorylation by aurora kinase A induces Mdm2-mediated destabilizationand inhibition of p53” Nature Genet. 36:55-62 (2004) PMID: 14702041
2. Blaydes JP et al “Stoichiometric phosphorylation of human p53 at Ser315 stimulates p53dependent transcription”J Biol Chem 276:4699-4708 (2001) PMID: 11078726
3. Bode AM & Dong Z “Post-translational modification of p53 in tumorigenesis” Nature Rev Cancer
4: 793-805 (2004) PMID: 15510160
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BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/