View the slide viewer

Have questions about this assay kit?

Human p53 Assay Kit

2x-48 assays in-96 well assay plates
2x-48 assays in-96 well assay plates

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Tumor Protein p53 (p53). INDIGO’s p53 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the Human Tumor Protein p53. In addition to p53 Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference activator, Luciferase Detection Reagent, and two cell culture-ready assay plates. The principal application of this reporter assay is in the screening of test compounds to quantify any functional activity that they may exert against p53. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.


  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeTranscription Factor
Receptor FormNative
Assay ModeAgonist
Kit Components
  • P53 Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Doxorubicin (hydrochloride), (ref. activator p53; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Orthologs AvailableNo
Shipping RequirementsDry Ice
Storage temperature-80C


Activation of p53 is demonstrated by treating reporter cells with Doxorubicin hydrochloride (provided), Quinacrine hydrochloride hydrate, (±)-Nutlin-3, Tenovin-1 (all from Cayman Chemical), and Cisplatin (Tocris). Reporter Cells were treated following the ‘~24 hour + ~24 hour’ dosing regimen described in the Technical Manual. Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n = 3). Fold-activation and Z’ values were calculated as described by Zhang, et al. (1999). All graphical manipulations were performed using GraphPad Prism software. The high Z' score confirms the robust performance of this assay.

Target Background

Tumor protein P53, also known as p53, acquired its reputation as “guardian of the genome” through its ability to sense and respond to cellular stress, preventing accumulation of DNA damage and subsequent formation of malignancies. p53 is a transcription factor and tumor suppressor that responds to cellular stress by regulating genes involved in a diverse array of cellular responses, including but not limited to, cell cycle arrest, DNA repair, apoptosis, senescence, and autophagy, thus minimizing the negative consequences of genetic mutation. p53 is expressed at low levels in cells in the absence of stress, regulated by various factors including MDM2 (also known as HDM2), which acts as a negative regulator through its E3 ubiquitin ligase activity, and WIP1/PPM1D, which also acts as a negative regulator of p53 through its ability to dephosphorylate specific residues on both p53 and MDM2, leading to the destabilization of p53 and the stabilization of HDM2, respectively. p53 in turn directly influences the expression of these two negative regulators, as both MDM2 and WIP1/PPM1D are target genes of p53.

p53 is the most commonly mutated gene in human cancer formation, and is therefore of interest in cancer research and therapeutic development. Potential therapeutics may target the p53 pathway in several ways. These include restoration of mutated p53 to its wild-type conformation and targeting regulators of p53 activity, for example, by inhibiting MDM2. The small molecule PRIMA-1, as well as derivates of the thiosemicarbazone family, have been shown to restore wild-type p53 activity, and several classes of MDM2 inhibitors, including nutlins, have been developed and studied. Since the causes, results, and regulation of p53 activation are diverse and context-dependent, therapeutics targeting the p53 pathway are necessarily diverse as well.

INDIGO’s p53 Reporter Cells include the luciferase reporter gene functionally linked to a p53-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in human p53 activity.

Therefore, the principal application of this reporter assay is in the screening of test compounds to quantify any functional activity that they may exert against human p53.

Also available as a service

Tumor Protein 53 (p53)

Have questions about this assay kit?