Invitrogen™ Human p53 ELISA Kit

Description

Includes

Aluminium pouch(es) with a Microwell Plate coated with monoclonal antibody to human p53
Biotin-Conjugate anti-human p53 monoclonal antibody
Streptavidin-HRP
Human p53 Standard lyophilized, 100 U/ml upon reconstitution
Sample Diluent
Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
Wash Buffer Concentrate 20x (PBS with 1% Tween 20)
Substrate Solution (tetramethyl-benzidine)
Stop Solution (1M Phosphoric acid)
Blue-Dye
Green-Dye
Red-Dye
Adhesive Films

ELISA

The Human p53 ELISA quantitates Hu p53 in human serum or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu p53. Principle of the method The Human p53 solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen. 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
• Assay Range: 0.78 to 50 U/mL
• Assay Sensitivity: 0.33 U/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Serum, Supernatant
• Target Species: Human
• For Use With (Application): ELISA
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 7157
• Gene Symbol: BCC7,LFS1,P53,TRP53
• Interassay CV: 8.9%
• Intraassay CV: 5.5
• Kit Contents: Pre-coated 96 well plate, Standard, Sample Diluent, Assay Buffer concentrate, Biotinylated Detection Antibody, SAV-HRP, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Serum 50 μL, Supernatant 50 μL
• Storage Requirements: 2°C to 8°C
• Synonym: Tumor suppressor p53, TP53, Li-Fraumeni syndrome
• Target: p53
• Test Time: 1 hrs 20 mins
• Total Assay Time: 3 hrs 10 mins