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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 | LC3B |
|---|---|
| Assay Target Class | Protein |
| Automation Compatible | Yes |
| Detection Method | Time-Resolved Fluorescence (TRF), TR-FRET |
| Experimental Type | In vitro |
| Shipping Condition | Blue Ice |
| Therapeutic Area | Autophagy |
| Unit Size | 500 Assay Points |
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
