Revvity, 64ARV7PEG, HTRF Androgen Receptor (Variant 7/AR-V7) Detection Kit, 500 Assay Points

Overview
The HTRF Androgen Receptor Variant 7 detection assay monitors the expression of the Androgen Receptor splice Variant 7 (AR-V7), providing insights into its presence, whether endogenous or overexpressed, within various cells.
The Androgen Receptor is a nuclear receptor that is activated by androgen hormones, including testosterone (T) and Dihydrotestosterone (DHT). Androgens bind to the ligand binding domain of the Androgen Receptor, resulting in receptor homodimerization and translocation to the nucleus. Downstream gene expression and signaling pathways are then activated, including the PI3K/AKT pathway. The gene expression induced by the Androgen Receptor promotes cell division, proliferation, differentiation, apoptosis, and angiogenesis. Dysregulation of the Androgen Receptor pathway can lead to oncogenic phenotypes, with Androgen Receptor activation or subexpression having been linked to conditions such as prostate cancer, breast cancer, glioblastoma, and other malignancies. Moreover, Androgen Receptor variants are expressed in various human cancer cell lines.
The Androgen Receptor Variant 7 (AR-V7) is a splice variant of the androgen receptor mRNA characterized by a truncated ligand-binding domain. It is constitutively active and widely expressed in human cancer cell lines. Among the AR-Vs described to date, AR-V7 is the most extensively evaluated and characterized, and several blood-based tests for AR-V7 have been developed.
Androgen Receptor and Variant 7 in particular has therefore become an important drug target for the pharmaceutical industry. The development of PROTAC compounds targeting the degradation of the Androgen Receptor is a popular therapeutic strategy to target Androgen Receptor-positive cancers.
How it works
HTRF androgen receptor variant 7 assay principle

The HTRF Human Androgen Receptor Variant 7 assay quantifies the expression level of Androgen Receptor in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Human Androgen Receptor Variant 7 assay uses two labeled antibodies, one coupled to a donor fluorophore and the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of Androgen Receptor 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 androgen receptor variant 7 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 Androgen Receptor Variant 7 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

Human Androgen Receptor Variant 7 one-plate assay protocol

Detection of Human Androgen Receptor Variant 7 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-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Technology-TR-FRET
Unit Size-500 assay points