R&D Systems社の抗p53抗体 抗p53抗体

※ 本製品は研究用です。研究用以外には使用できません。

R&D Systems社の抗p53抗体です。各種アプリケーションに使用可能な製品が揃っています。

Molecule Information: p53

Background: p53

p53 is well known for its key role as a tumor suppressor protein. It is 393 amino acids (aa) in length with a predicted molecular weight of 44 kDa. It belongs to the p53 family that also includes p63 and p73. Structurally, p53 is characterized by an N-terminal transactivation domain, central DNA-binding and oligomerization domains, and a C-terminal regulatory domain. It is thought to exist as a homotetramer, and it exhibits approximately 72% and 76% aa identity with its mouse and rat orthologs, respectively. Mutations in the p53 gene are one of the most frequent genomic events accompanying oncogenic transformation. p53 responds to signals such as DNA damage or cell stress primarily through its actions as a transcription factor. Among its gene targets are a range factors that promote DNA repair mechanisms or apoptosis including cell cycle regulatory proteins and members the Bcl-2 family. Because of its critical role in genomic homeostasis, p53 activities are tightly regulated by a network of protein-protein interactions, microRNAs, and a range of post-translational modifications, including phosphorylation, acetylation, methylation, and ubiquitination. A widely studied regulator is Murine Double Minute 2 (MDM2). MDM2 is known to suppress p53 activity through direct binding or through its actions as a Ubiquitin ligase (E3) that catalyzes p53 ubiquitination and proteasome-mediated degradation.

Application & Data: Human Phospho-p53 (S46) Polyclonal Antibody (#AF1489 )

■Western Blot

Detection of Human Phospho-p53 (S46) by Western Blot.
Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 1 µM camptothecin (CPT) for 5 hours. PVDF membrane was probed with 1 µg/ml Rabbit Anti-Human Phospho-p53 (S46) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1489) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-p53 (S46) was detected at approximately 53 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda -phosphatase ( lambda -PPase) for 30 minutes. For additional reference the membrane was stripped and reprobed for total p53 using Human/Mouse/Rat p53 HRP-conjugated Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # HAF1355). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

■Immunohistochemistry

p53 in Human Breast Cancer Tissue.
p53 phosphorylated at S46 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-p53 (S46) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1489) at 1.7 µg/ml overnight at 4℃. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue).