Revvity AlphaLISA Human and Mouse LC3B-II Detection Kit, 5,000 assay points

LC3 is a mammalian homolog of the yeast ATG8 protein and is distributed throughout mammalian tissues and cultured cells. LC3 is a key component of the autophagy process - a degradative program that maintains cellular homeostasis. During autophagy, cytoplasmic components such as cytosolic proteins and organelles are engulfed by autophagosomes Concurrently, a cytosolic form of LC3 (LC3-I) is conjugated to phosphatidylethanolamine to form LC3-phosphatidylethanolamine conjugate (LC3-II). This newly formed conjugate is recruited to autophagosomal membranes. The subsequent fusion of autophagosomes with lysosomes results in the formation of autolysosomes, and intra-autophagosomal components are degraded by lysosomal hydrolases. At the same time, LC3-II within the autolysosomal lumen is degraded. The LC3-II level has been found to be proportional to autophagosome abundance making LC3-II the most popular marker of autophagy.

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

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.