Western blot Biotin

Get tips on using RNeasy PowerBiofilm Kit (50) to perform RNA isolation / purification Biofilm

Get tips on using DNeasy PowerBiofilm Kit (50) to perform DNA isolation / purification Biofilm samples

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.

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.

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 Investigator Quantiplex HYres Kit to perform DNA quantification Blood

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - immortalized Human blood cord

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human peripheral blood monocytes

Get tips on using QIAamp DSP Virus Kit to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary canine peripheral blood mononuclear cells