Background: Autophagy is a lysosomal degradation pathway for cytoplasmic material, which is activated during stress conditions such as amino acid starvation or viral infection. Mammalian cells use autophagy during short periods of starvation to degrade nonessential cellular components in order to liberate nutrients for vital biosynthetic reactions. Recent results have shown that autophagy also contributes to development, growth regulation and cancer, as well as longevity. After induction by a stress signal such as amino acid starvation, the first step in autophagy is the formation of an autophagosome. A well published autophagosome marker protein, MAP LC3, was originally identified as a microtubule associated protein and named 'microtubule-associated-protein-light-chain-3'. LC3 is a small 16-18 kDa protein that is soluble in nonstarved cells, but becomes peripherally membrane-associated during amino acid starvation. By immunoelectron microscopy, LC3 has been shown to associate to the inner and outer limiting membranes of autophagosomes, and the membrane association is mediated by a covalent conjugation to a lipid, phosphatidylethanolamine. In Western blots, two forms of LC3 are seen, LC3-I and LC3-II. LC3-I is found in the soluble fraction, and LC3-II in the pelletable membrane fraction. Both LC3-I and LC3-II are seen in nonstarved cells, but during autophagy induction the proportion of LC3-II increases. Autophagy ELISA Kit (LC3-II Quantitation) is an enzyme immunoassay developed for detection and quantitation of LC3 in cell lysate samples. The kit utilizes a selective permeabilization procedure to remove the cytosolic pro-LC3 and LC3-I and retain the autophagosome membrane bound LC3-II (Figure 1). The quantity of LC3-II in unknown lysate sample is determined by comparing its absorbance with that of a known LC3 standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

Small volumes of Autophagy elisa kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS169564 is a ready-to-use microwell, strip-or-full plate ELISA (enzyme-linked immunosorbent assay) Kit for analyzing the presence of the Autophagy, ELISA Kit target analytes in biological samples. The concentration gradients of the kit standards or positive controls render a theoretical kit detection range in biological research samples containing Autophagy. The ELISA analytical biochemical technique of the MBS169564 kit is based on Autophagy antibody-Autophagy antigen interactions (immunosorbency) and an HRP colorimetric detection system to detect Autophagy antigen targets in samples. The ELISA Kit is designed to detect native, not recombinant, Autophagy. Appropriate sample types may include undiluted body fluids and/or tissue homogenates, secretions. Quality control assays assessing reproducibility identified the intra-assay CV (%) and inter-assay CV(%).