Revvity, AL3209C, AlphaLISA Human and Mouse α-SMA Detection Kit, 500 Assay Points

Overview
Alpha-Smooth Muscle Actin or α-SMA is a member of Actin family that forms microfilaments and is used as a marker of myofibroblast formation. 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.
Formats:

Our 100 assay point kit allows you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).

Features:

No-wash steps, no separation steps
ELISA alternative technology
Sensitive detection
Broad sample compatibility
Small sample volume
Results in less than 4 hours

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
How it works
Principle of the AlphaLISA human & mouse α-SMA assay
The AlphaLISA α-SMA assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-target antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-target antibody conjugated to AlphaLISA Acceptor beads. Two antibodies are directed against the α-SMA protein. In the presence of the target, both antibodies bind to α-SMA and the beads come into proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer within the Acceptor beads, resulting in emission with λmax at 615 nm. The intensity of the signal is directly proportional to the concentration of α-SMA present in the sample (cell lysate).
 

Protocol of the AlphaLISA human & mouse α-SMA assay
The AlphaLISA α-SMA assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples (cell lysates) or control lysate are dispensed directly into the assay plate for the detection of α-SMA by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
Application-Protein Quantification
Automation Compatible-Yes
Brand-AlphaLISA
Detection Modality-Alpha
Protocol Time-3h at RT
Sample Volume-5 µL
Shipping Conditions-Shipped in Blue Ice
Target-α-SMA
Target Class-Biomarkers
Target Species-Human Mouse
Technology-Alpha
Therapeutic Area-Cardiovascular NASH/Fibrosis
Unit Size-500 assay points