Revvity, 64HPRTPEH, HTRF Human Total Progesterone Detection Kit, 10,000 Assay Points

Overview
The HTRF Human Progesterone Receptor detection assay monitors the expression of endogenous or overexpressed Progesterone Receptor in various cells. The kit recognizes PRA and PRB isoforms.
Progesterone Receptor (PR) is a nuclear receptor that is activated by progesterone. The hormone crosses the cell membrane and binds to the receptor. After binding, PR homodimerizes, enabling translocation to the nucleus. Coupled with a co-activator, the receptor dimer will be recruited to the Progesterone Response Element (PRE) and induce gene expression leading to proliferation / apoptosis balance. PR expression is modulated by Estrogen Receptor. Therefore, PR is a biomarker commonly used as an indicator of Estrogen Receptor-α function. The MAPK kinase pathway has also been described as having an effect on PR activity through phosphorylation.
How it works
Human Total Progesterone Receptor Detection Kit assay principle
The HTRF Human Progesterone Receptor assay quantifies the expression level of Progesterone Receptor in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Human Progesterone 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 the presence of Progesterone 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 Total Progesterone Receptor 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 Progesterone Receptor HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Human Total Progesterone Receptor Detection Kit one-plate assay protocol
Detection of Human Progesterone 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
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Unit Size-10,000 assay points