An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples 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 Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCAEC
Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCtASMC
Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCtAEC
Get tips on using ProcartaPlex Human Cytokine Panel 1B, 25 plex to perform ELISA (kit) Human Serum Cytokine measurements (Multiplex assay) - -NA- Human -NA-
Get tips on using Senescence Detection Kit - Merck to perform Reporter gene assay β-galactosidase substrates - MCF-7 human breast cancer
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 CpGenome Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - Human ovarian tissue TGFBI
Get tips on using GenElute™ Mammalian Total RNA Miniprep Kit to perform RNA isolation / purification Cells - primary human pancreatic stellate cells
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