As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.
ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.
Get tips on using Tissue or Cell Total Protein Extraction Kit to perform Protein isolation Tissue - Lamb muscle tissue
Get tips on using DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit to perform ROS assay cell type - L-02 human fetal hepatocyte
Get tips on using miRNeasy FFPE Kit to perform RNA isolation / purification Tissue - Human Kidney
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Tissue - Human Veins
Get tips on using miRNeasy FFPE Kit to perform RNA isolation / purification Tissue - Human Tonsil
Get tips on using miRNeasy FFPE Kit to perform RNA isolation / purification Tissue - Human Prostate
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Tissue - Human Muscles
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Tissue - Human Larynx
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