rna-isolation-purification-cells-primary-human-carotid-artery-endothelial-cells

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Tissue - Rat Adrenal glands

Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Tissue - rat heart tissue

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 LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - mouse bone marrow-derived macrophages

Get tips on using RNAprep pure Kit (For Plant) to perform RNA isolation / purification Yeast - Neurospora crassa

Get tips on using Galacto-Star™ β-Galactosidase Reporter Gene Assay System for Mammalian Cells 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.

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