Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical artery smooth muscle cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human pulmonary artery smooth muscle cells
Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human pulmonary artery smooth muscle cells
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.
TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.
Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary porcine coronary artery endothelial cells
Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human coronary artery endothelial cells
Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary porcine coronary artery endothelial cells
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human coronary artery endothelial cells
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