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The LANCE® Ultra Human LC3B Detection Kit is designed for detection and quantitation of human LC3B in cell culture media using a homogeneous TR-FRET (no-wash steps, no separation steps) assay.
LANCE® and LANCE® (Lanthanide chelate excite) Ultra are our TR-FRET (time-resolved fluorescence resonance energy transfer), homogeneous (no wash) technologies. One antibody of interest is labeled with a donor fluorophore (a LANCE Europium chelate) and the second molecule is labeled with an acceptor fluorophore (ULight™ dye). Upon excitation at 320 or 340 nm, energy can be transferred from the donor Europium chelate to the acceptor fluorophore if sufficiently close for FRET (~10 nm). This results in the emission of light at 665 nm.
LC3 represents a mammalian homologue of the yeast autophagy related gene ATG8. It was originally characterized as light chain 3 of the microtubule associated protein 1 (MAP1LC3). The protein family consists of LC3 A, B, and C and the GABARAP subfamily. Human LC3B is 125 amino acids long. After synthesis, it is cleaved by ATG4B to expose a Cterminal glycine, representing the cytosolic form LC3B I. During autophagy the C-terminus is covalently linked to autophagosomal vesicle membranes via a phospholipid anchor and this form is called LC3B II. The transformation of LC3B I to II is mediated by a ubiquitination-like process involving ATG7 (E1), ATG3 (E2) and the ATG16L complex (E3). To date, LC3B is considered as the most persistent marker of the autophagy pathway.
|Assay Target Class||Protein|
|Detection Method||Time-Resolved Fluorescence (TRF), TR-FRET|
|Experimental Type||In vitro|
|Shipping Condition||Blue Ice|
|Unit Size||10,000 Assay Points|