siRNA / RNAi /miRNA transfection Bovine

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.

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 TIANamp Genomic DNA Kit to perform DNA isolation / purification Tissue - bone

Get tips on using QIAamp DNA Mini Kit to perform DNA isolation / purification Tissue - bone

Get tips on using QIAamp DNA FFPE Tissue Kit to perform DNA isolation / purification Tissue - bone

Get tips on using In Vitro Angiogenesis Assay Kit to perform Angiogenesis assay human - bone marrow mononuclear cells

Get tips on using Minimum Essential Medium Eagle to perform Stem cell culture media Human bone mesenchymal stem cell (BMSC)

Get tips on using Atg5 (D5F5U) Rabbit mAb to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)

Get tips on using Anti-LC3B antibody (ab48394) to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)