shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y) Connexin 43

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - human peripheral blood mononuclear cells (PBMCs)

Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - human colorectal adenocarcinoma cells (CL-187)

Get tips on using RNAqueous™ Total RNA Isolation Kit to perform RNA isolation / purification Cells - primary human lung fibroblasts

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human osteoblasts

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

Get tips on using ARCTURUS® PicoPure® DNA Extraction Kit to perform RNA isolation / purification Cells - primary human melanocytes

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.

Get tips on using Quant-iT™ RiboGreen™ RNA Assay Kit to perform RNA quantification Fuorimetric - human trophoblast cells