Revvity, AL3199F, AlphaLISA Human NFL Detection Kit, 5,000 Assay Points

Overview
Neurofilament Light chain protein, also known as Neurofilament Light polypeptide, NF-L, NFL or NEFL is a member of intermediate filament protein family, selectively expressed in neurons. NF-L (68 kDa) associated with neurofilament-heavy chain (NF-H, 200 kDa), medium chain (NF-M, 125 kDa) and alpha-internexin, forms the structure of axons. Depending on conditions, different levels of NF-L are released during brain development, maturation, aging and in numerous neurodegenerative diseases proportionally to the degree of axonal damage. Many studies have demonstrated that NF-L can be used as a biomarker for disease monitoring and more broadly as a neuronal damage marker but also to check the differentiation into neurons in IPSc models for example.
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 3 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 Neurofilament Light chain assay
The AlphaLISA Neurofilament Light chain (NF-L) assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-hNF-L antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-NF-L antibody conjugated to AlphaLISA Acceptor beads. Two antibodies are directed against the hNF-L protein. In the presence of the hNF-L, both antibodies bind to hNF-L 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 hNF-L present in the sample (cell supernatant).

Protocol of the AlphaLISA human Neurofilament Light chain assay
The AlphaLISA Neurofilament Light chain (NF-L) assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples (cell supernatants) or standards are dispensed directly into the assay plate for the detection of human NF-L 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
Host Species-Human
Product Group-Kit
Shipping Conditions-Shipped in Blue Ice
Target-Neurofilament L
Target Class-Cytokines
Target Species-Human
Technology-Alpha
Therapeutic Area-Neuroscience
Unit Size-5,000 assay points