Revvity, 62ASMAPEH, HTRF Human α-SMA Detection Kit, 10,000 Assay Points

Overview
After tissue injury, TGF-beta locally released by inflammatory cells activates resident fibroblasts or quiescent HSCs (hepatic stellate cells). This leads to their differentiation into myofibroblasts, whose role is to migrate into the damaged tissue and synthesize ECM (extracellular matrix) components to repair the wound. Myofibroblasts are characterized by de novo expression of alpha-SMA, which is incorporated into actin stress fibers and confers a high contractile activity to the cells. Chronic tissue injury leads to persistent de novo formation of myofibroblasts (alpha-SMA+), excessive contraction, and deposition of ECM, eventually leading to tissue fibrosis.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
Assay principle
The alpha-SMA assay is based on a TR-FRET sandwich immunoassay format comprising two antibodies, one labelled with Eu3+-Cryptate (donor) and the other with d2 (acceptor). The antibodies specifically bind to Alpha-SMA, and the proximity of donor and acceptor leads to a fluorescent TR-FRET signal.

Assay protocol
Two-plate assay protocol For added flexibility, the assay can be run under a two-plate assay protocol, where cells are plated, treated, and lysed in a culture plate. For detection, lysates are subsequently transferred into a 96- or 384-well low volume detection microplate, where the HTRF reagents are added. This protocol enables the cells' viability and confluence to be monitored. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. The assay can be run with frozen cell lysates or fresh cells in culture. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Biomarkers
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Cardiovascular NASH/Fibrosis
Unit Size-10,000 assay points