siRNA / RNAi /miRNA transfection Human Cells THP-1

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.

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.

Get tips on using miRNeasy Serum/Plasma Kit to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

Get tips on using pcDNA3.1-MRP4-his6 to perform Protein Expression Eukaryotic cells - HEK293 human MRP4-his6

Get tips on using pET-28b-hBCO1 to perform Protein Expression Prokaryotic cells - E. coli human BCO1

Get tips on using pGL3-Basic Vector to perform Reporter gene assay luciferase - human embryonic stem cells

Get tips on using IGEPAL® CA-630 to perform Protein isolation Mammalian cells - Human lung fibroblasts

Get tips on using NanoOrange™ Protein Quantitation Kit to perform Protein quantification Mammalian cells - Human podocytes

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.