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.
Get tips on using CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) to perform
Get tips on using LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit to perform Live / Dead assay mammalian cells - mouse keratinocytes
Get tips on using Flp-In™ T-REx™ 293 Cell Line to perform Protein expression and purification Mammalian cells - HeLa ChaC1
Get tips on using EZViable™ Calcein AM Cell Viability Assay Kit (Fluorometric) to perform Live / Dead assay mammalian cells - rat brain microvascular endothelial cells
Get tips on using Flp-In™ T-REx™ 293 Cell Line to perform Protein expression and purification Mammalian cells - CAL-51 BRCA1
Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,
Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment