Invitrogen™ p53 Multispecies In-Cell ELISA Kit, Colorimetric

Product Code. 11548110

Brand: Invitrogen™ 62216

Description

In-cell ELISA Kit

The p53 Multispecies In-Cell ELISA Colorimetric Detection Kit is a simple method for quantifying intracellular proteins in whole cells. Principle of the method To perform the assay, cells are first plated, treated and fixed. Expression of the protein(s) of interest is monitored in wells of a microplate using target-specific primary antibodies (see the Important Product Information section for antibodies included in each kit) and a horseradish peroxidase (HRP)-conjugated detection reagent. The kit is supplied with a whole-cell stain to control for differences in cell plating, which is important when measuring relative levels of a protein with different treatments or assessing its post-translational modification (PTM) form. After staining, the results are analyzed by normalizing the absorbance (HRP activity) values to cell number, which adjusts for the cell plating differences among the wells. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

The tumor suppressor protein, p53, is a sequence specific transcription factor that is activated by cellular stress. p53 mediates cell cycle arrest or apoptosis in response to DNA damage or starvation for pyrimidine nucleotides. p53 is up-regulated in response to stress signals and stimulated to activate transcription of specific genes, resulting in expression of p21waf1 and other proteins involved in G1 or G2/M arrest. The structure of p53 comprises an N-terminal transactivation domain, a central DNA-binding domain, an oligomerisation domain, and a C-terminal regulatory domain. There are various phosphorylation sites on p53, of which the phosphorylation at Ser15 is important for p53 activation and stabilization. p53 has been characterized to play a role in blocking the proliferative action of damaged cells and act as an anticancer agent. Phosphorylation of Ser392 in p53 has been shown to associate with the formation of human tumors. In addition, p53 has also been linked to the effects of aging and oxidative stress and an increase in p53 has been linked to deficits in LTP (Long Term Potentiation) in learning and memory. p53 is found in very low levels in normal cells, however, in a variety of transformed cell lines, it is expressed in high amounts, and believed to contribute to transformation and malignancy. Mutants of p53 that frequently occur in a number of different human cancers fail to bind the consensus DNA binding site, and cause the loss of tumor suppressor activity. Alterations of the TP53 gene occur not only as somatic mutations in human malignancies, but also as germline mutations in some cancer-prone families such as Li-Fraumeni syndrome.

Specifications

• Accession Number: P04637
• Description: Pierce p53 Colorimetric In-Cell ELISA Kit
• Conjugate: HRP
• Immunoassay Kit Format: Multi-component kit with HRP-conjugate
• Product Type: ELISA Kit
• Sample Type: Supernatant
• Sufficient For: 96 microplate wells of overnight-cultured cells
• Target Species: Human
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 7157
• Gene Symbol: BCC7,LFS1,P53,TRP53
• Kit Contents: Blocking Buffer, 20X Tris Buffered Saline, Surfact-Amps 20, Surfact-Amps X-100, HRP Conjugate, TMB Substrate, TMB Stop Solution, Janus Green Whole-Cell Stain, Elution Buffer, Thin Plate Seal Assembly, Components included only in the target-specific kits:Antibody #1 and Antibody #2, Janus Green Whole-Cell Stain, 50mL
• Label or Dye: HRP
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Supernatant, 10,000 cells
• Storage Requirements: 2°C to 8°C
• Target: Human
• Test Time: 1 hr. 20 min.
• Total Assay Time: 24 hr.