It accumulates inside the acidic parts of the cell, including endosomes and lysosomes. This accumulation leads to inhibition of lysosomal enzymes that require an acidic p H, and prevents fusion of endosomes and lysosomes. TLR7/8-Mediated Activation of Human NK Cells Results in Accessory Cell-Dependent IFN- Production. G6pd and hydroxychloroquine Can hydroxychloroquine affect memory For example, starvation-induced autophagy has been confirmed by a myriad of different assays and can also be clearly observed with an autophagic flux assay via an LC3 western blot. To perform this assay which can easily be adapted to any stimulus, four conditions should be analyzed. Inhibition of autophagy by administration of chloroquine CQ in combination anticancer therapies is currently evaluated in clinical trials. We observed in 3 different human cancer cell lines cultured at acidic pH that autophagic flux is not blocked by CQ. The Autophagy Flux Detection kit facilitates the simple measurement of autophagy in cells. The kit utilises highly characterised antibodies to key autophagy biomarkers, LC3-II and p62, together with application of specific high purity control inhibitor for the accurate assessment of autophagic flux by Western blotting methods. Moreover, Chloroquine inhibits autophagy as it raises the lysosomal p H, which leads to inhibition of both fusion of autophagosome with lysosome and lysosomal protein degradation . Chloroquine is commonly used to study the role of endosomal acidification in cellular processes [2, 3], such as the signaling of intracellular TLRs. Chloroquine autophagy flux Autophagic Flux, Fusion Dynamics, and Cell Death., Chloroquine & Hydroxychloroquine supporting chemo. Drug reaction with acyclovir and sucralfate and hydroxychloroquineRetinal thinning in plaquenil toxicity octChloroquine resistance treatmentChloroquine treatment for vivax malaria LC3-II Autophagy Enrichment Kits. LC3 precursors are proteolytically processed to form LC3-I, which is diffusely distributed in the cytosol. Upon initiation of autophagy, the C-terminal glycine of LC3-I is modified by addition of a phosphatidylethanolamine PE to form LC3-II, which translocates rapidly to nascent autophagosomes in a punctate distribution 5. Autophagy Assays Sigma-Aldrich. Autophagy Flux Detection Kit StressXpress® SKT-135.. Autophagic flux determines cell death and survival in.. Autophagy pathway and target inhibitors Combination of autophagy related Antibodies and Inhibitors MBL provides a useful set of antibodies and inhibitors that can detect autophagy flux! Anti-LC3 antibodies × 2 items Loading control antibody Positive control for LC3 Inhibitors × 2 items Cell lysis buffer Bafilomycin A1 V type ATPase specific inhibitor SQSTM1 encodes the cargo adaptor protein, p62, which interacts with autophagic substrates and delivers them to autophagosomes for degradation. In the process, p62 is itself degraded and when autophagy is induced, a corresponding decrease in p62 levels is observed. Chloroquine is a lysosomotropic agent that prevents endosomal acidification 1. It accumulates inside the acidic parts of the cell, including endosomes and lysosomes. This accumulation leads to inhibition of lysosomal enzymes that require an acidic pH, and prevents fusion of endosomes and lysosomes.