Revvity, 64ANDRPEH, HTRF Human Androgen Receptor Detection Kit, 10,000 Assay Points

Overview
The HTRF Human Androgen Receptor detection assay monitors Wild-Type Androgen Receptor (AR-WT) and the Androgen Receptor splice Variant 7 (AR-V7), and is used to detect the expression of endogenous or overexpressed Androgen Receptors in various cells.
Androgen Receptor is a type of nuclear receptor that is activated by androgen hormones, like 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. This activates downstream gene expression and activation of signaling pathways, like the PI3K/AKT pathway. The gene expression induced by Androgen Receptor results in cell division, proliferation, differentiation, apoptosis, and angiogenesis. A dysregulation of the Androgen Receptor pathway could lead to oncogenic phenotypes, and Androgen Receptor activation or surexpression has been widely linked to prostate cancer as well as other types of cancer, like breast cancer and glioblastoma. Moreover, Androgen Receptor variants are expressed in various human cancer cell lines. The Androgen Receptor Variant 7 (AR-V7), a splice variant of the androgen receptor mRNA resulting in the truncation of the ligand-binding domain, is constitutively active and widely expressed in human cancer cell lines. Androgen Receptor has therefore become an important drug target for the pharmaceutical industry. The development of PROTAC compounds to target Androgen Receptor degradation is a part of the therapeutic strategy to target Androgen Receptor positive cancers.
How it works
HTRF Human androgen receptor assay principle
The HTRF Human Androgen Receptor 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 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 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 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 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Human androgen receptor one-plate assay protocol
Detection of Human Androgen Receptor 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
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 2 Lysis Buffer 3
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points