siRNA / RNAi /miRNA transfection Human Cells OV-2008

Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using KnockOut™ Serum Replacement - Multi-Species to perform Stem cell Differentiation media Human Limbal Epithelial cells

Get tips on using Anti-LC3 antibody produced in rabbit to perform Autophagy assay cell type - Human osteosarcoma cancer cells

Get tips on using PE Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - Human T-cells

Get tips on using FITC Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - Human T-cells

Get tips on using MethylCap kit to perform DNA methylation profiling Whole genome profiling - HCT116, HTC15 human colon cancer cells

Get tips on using M-PER™ Mammalian Protein Extraction Reagent to perform Protein isolation Tissue - Human aortic endothelial cells

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.

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.

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.