protein-expression-and-purification-mammalian-cells-hek-293-mt-pa

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 Wizard® Plus Midipreps DNA Purification System Technical Bulletin to perform Plasmid Isolation Proteus mirabilis

Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 ROCK2

Get tips on using N788 phenol-free total RNA purification kit to perform RNA isolation / purification Bacteria - Gram negative Pseudomonas aeruginosa

Get tips on using Cell Cycle and Apoptosis Analysis Kit to perform Cell cycle assay human - Hep G2

Get tips on using NeuroMag to perform DNA transfection Mammalian cells - Immortalized cell lines SH-SY5Y

Get tips on using Cell Counting Kit-8 to perform Live / Dead assay mammalian cells - GH3

Get tips on using EZ4U - Cell Proliferation Assay to perform Live / Dead assay mammalian cells - GH3

Get tips on using Minimum Essential Medium Eagle to perform Mammalian cell culture media HSG cells

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.