• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Assay Length

  4 hours 40 mins (after plate preparation)

• Sample Type & Volume Required

  Cell lysates (100 µL)

• Range

  125.00 - 8,000 pg/mL

• Sufficient Materials

  Kits available for two, five, or fifteen 96-well plates*

• Specificity

  Please see the

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Product Features

• Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time

• Development protocols are provided to guide further assay optimization

• Assay can be customized to your specific needs

• Available in 2, 5, and 15- (96-well) plate pack sizes

• Economical alternative to Western blot

Kit Content

• Capture Antibody

• Conjugated Detection Antibody

• Calibrated Immunoassay Standard or Control

• Streptavidin-HRP

Other Reagents Required

  PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na  ₂HPO  ₄, 1.5 mM KH  ₂O  ₄, pH 7.2 - 7.4, 0.2 µm filtered  
 
  Wash Buffer: (Catalog # ), or equivalent  
 
  Lysis Buffer*  
 
  IC Diluent*

Blocking Buffer*  
 
  Substrate Solution: 1:1 mixture of Color Reagent A (H  ₂O  ₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # )  
 
  Stop Solution: 2 N H  ₂SO  ₄ (Catalog # )  
 
  Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # ), or equivalent  
 
  Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent  
 

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Preparation and Storage

• Storage

  Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

• Entrez Gene IDs:

  7157 (Human); 22059 (Mouse); 24842 (Rat);

• Aliases:

  Antigen NY-CO-13; BCC7; FLJ92943; LFS1; LFS1TRP53; p53 tumor suppressor; p53; P53cellular tumor antigen p53; Phosphoprotein p53; TP53; transformation-related protein 53; TRP53; tumor protein p53; Tumor suppressor p53