siRNA / miRNA gene silencing Human hES cell line H1 (WA01)

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 OSTEOPONTIN (O-17) ANTI-HUMAN RABBIT IGG AFFINITY PURIFY to perform Immunohistochemistry Mouse - Spp1/OPN

Get tips on using Cell Comb™ Scratch Assay to perform Wound healing assay cell type - mouse 3T3-L1

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary human aortic smooth muscle cells

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Immortalized cell lines HEK293

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

Get tips on using PolyFect Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HEK293

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,

Get tips on using Human/Mouse/RatTotal SOD2/Mn-SOD DuoSet IC ELISA to perform ELISA Rat - SOD2/Mn-SOD

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.