rna-isolation-purification-cells-primary-rat-brain-microvascular-endothelial-cells

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.

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 RNeasy Mini Kit to perform RNA isolation / purification Cells - primary rat marrow stromal cells

Get tips on using Absolutely RNA Nanoprep Kit to perform RNA isolation / purification Cells - primary rat dorsal root ganglion neurons

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary rat retinal pigment epithelium cells

Get tips on using E.Z.N.A.® Total RNA Kit I to perform RNA isolation / purification Cells - primary rat cardiac fibroblasts

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary rat lymph node cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rat osteoblasts

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rat astrocytes

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary rat aortic smooth muscle cells