Revvity, 64ERAPEH, HTRF Human Total Estrogen Receptor α Detection Kit, 10,000 Assay Points

Overview
The HTRF Human Estrogen Receptor Alpha detection assay monitors the expression of endogenous or overexpressed Estrogen Receptor Alpha in various cells.
Estrogen Receptor Alpha is a type of nuclear receptor that is activated by estrogen hormones, like Estradiol (E2).
Estrogens bind to the ligand binding domain of the Estrogen Receptor Alpha, resulting in receptor homodimerization and translocation to the nucleus. This activates downstream gene expression and the activation of signaling pathways, like the PI3K/AKT pathway. The gene expression induced by Estrogen Receptor Alpha results in cell division and proliferation. A dysregulation of the Estrogen Receptor Alpha pathway could lead to oncogenic phenotypes, and Estrogen Receptor Alpha activation or overexpression has been widely linked to breast cancer (in more than 70% of the cases). Estrogen Receptor Alpha has therefore become an important drug target for the pharmaceutical industry. The development of PROTAC compounds to target Estrogen Receptor Alpha degradation is a part of the therapeutic strategy to target Estrogen Receptor Alpha positive cancers. Alternatively Selective Estrogen Receptor Degrader (SERD) can be detected with this kit, but not Selective Estrogen Receptor Modulator (SERM) as they only impact downstream signaling and not the expression level of the protein.
How it works
Human Total Estrogen Receptor alpha Detection Kit assay principle
The HTRF Human Estrogen Receptor-alpha assay quantifies the expression level of Estrogen Receptor-alpha in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Human Estrogen Receptor-alpha assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of Estrogen Receptor-alpha in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein's expression under a no-wash assay format

 

Human Total Estrogen Receptor alpha Detection Kit two -plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Human Estrogen Receptor-alpha HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Human Total Estrogen Receptor alpha Detection Kit one-plate assay protocol
Detection of Human Estrogen Receptor-alpha with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Protein Quantification
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Biomarkers
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points