Human Total p53 DuoSet IC ELISA Summary Assay Type | Solid Phase Sandwich ELISA | Format | 96-well strip plate | Assay Length | 4 hours 40 mins (after plate preparation) | Sample Type & Volume Required Per Well | Cell lysates (100 μL) | Sensitivity | 0.058 ng/mL | Assay Range | 125.0 - 8,000 pg/mL | Specificity | to measure total p53 in cell lysates | Cross-reactivity | < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested | Interference | No significant interference observed with available related molecules. |
Product Summary This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total p53 in cell lysates. An immobilized capture antibody specific for p53 binds both phosphorylated and unphosphorylated p53. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format. Preparation and Storage Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
Background: p53p53 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. Long Name: | transformation-related protein 53 | Entrez Gene IDs: | 7157 (Human); 22059 (Mouse); 24842 (Rat) | Alternate Names: | 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 |
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1、Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2、Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3、Add 50 μL of Assay Diluent to each well. 4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7、Aspirate and wash 5 times. 8、Add 100 μL Substrate Solution to each well. 9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
