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Invitrogen
Multispecies p53 In-Cell ELISA Kit, Colorimetric
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Product Specifications
Sample type/volume
Supernatant
10,000 cells
Hands-on time
1 hr 20 min
Time-to-result
24 hr
Homogenous (no wash)
No
Instrument
Absorbance Microplate Reader
Product size
96 Tests
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
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
Shipping conditions
Wet ice
Storage
2-8°C
Protein name
p53
Target site-specificity
Protein family
Other Proteins
Assay kit format
In-cell ELISA Kit
Detector antibody conjugate
HRP
Label or dye
HRP
About This 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.
Target information
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Bioinformatics
Protein Aliases : Antigen NY-CO-13, Phosphoprotein p53, Tumor suppressor p53
Performance Guarantee
If an Invitrogen™ immunoassay doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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