Phospho-p53 (Ser15) Translocation Assay Kit (Cell-Based) (BN01126)

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BN01126
  • Phospho-p53 (Ser15) Translocation Assay Kit (Cell-Based) (BN01126)
  • Phospho-p53 (Ser15) Translocation Assay Kit (Cell-Based) (BN01126)
  • Phospho-p53 (Ser15) Translocation Assay Kit (Cell-Based) (BN01126)
  • Phospho-p53 (Ser15) Translocation Assay Kit (Cell-Based) (BN01126)
$822

Description

ELISA Kit Technical ManualMSDS

Phospho-p53 (Ser15) Translocation Assay Kit (Cell-Based)

Tumor protein p53 (TP53, p53) is a widely conserved transcription factor and tumor suppressor commonly referred to as the "guardian of the genome". Due to the presence of transcriptional activation, DNA binding, and oligomerization domains, p53 responds to diverse cellular stresses to regulate expression of target genes, thereby inducing cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. Mutations in p53 gene are associated with numerous human cancers and about 50% of those mutations result in loss of function. Multiple genotoxic and non-genotoxic stimuli lead to stabilization and activation of p53 that are mediated by numerous post-translational modifications. The N-terminus of p53 is heavily phosphorylated, whereas the C-terminus may be phosphorylated, acetylated neddylated, ubiquitinated or sumoylated. In unstressed cells, the level of p53 protein is downregulated via binding to MDM2, COP1, PIRH2 or JNK that promote p53 degradation. Cellular stress and Nutlin-3, a potent inhibitor of Mdm2-p53 interaction, activate and increase level of p53 in the cell due to lack of negative regulation. Presence of three putative nuclear localization signals (NLSs) on its C-terminus, promotes translocation of some of the activated p53 into the nucleus thus activating genes that induce cell cycle arrest, senescence, or apoptosis. Assay Genie's Phospho-p53 (Ser15) Translocation Assay Kit provides a simple and complete assay in a ready to use format to visualize the nuclear translocation of activated p53 in mammalian cells.

Figure: Nutlin-3-induced activation and translocation of phosphorylated p53 in J774 cells. J774 cells were grown, fixed and stained according to the provided protocol. Panel A represents untreated cells whereas cells in panel B were treated for 4 hours with Nutlin-3. In the untreated cells, phosphorylated p53 is present in very low levels and localizes in the cytoplasm(A). Upon stimulation with Nutlin-3, phosphorylated p53 translocates into the nucleus which is reflected by strong nuclear fluorescence (B). Panel C shows activated cells in B counterstained with DAPI for DNA content. Panel D, a merger of B and C, demonstrates co-localization of phospho-53 and nuclear staining (magenta).

Key Information Description

Product SKU

BN01126

Size

50 assays

Detection Method

Fluorescence microscope capable of measuring EX at 570 nm and equipped with UV filter for DAPI

Species Reactivity

Human, mouse, rat

Applications

Detection of nuclear translocation of phosphorylated p53 in mammalian cells

Features and Benefits

  • Detection of nuclear translocation of phosphorylated p53 in mammalian cells
  • Characterize/Screen/Study for effectors of p53 cellular levels
  • Detection of cellular stress or DNA damage caused by genotoxic and non-genotoxic stimuli

Kit Components

  • Fixative Solution Blocking Buffer
  • Wash Buffer
  • Phospho-p53 Primary Antibody (100X)
  • Secondary Antibody (100X)
  • Nutlin-3
  • DAPI (1000X)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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