RNA isolation / purification Cells Cancer cell lines Leukemia cancer cell lines

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.

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 CellTiter-Glo® Luminescent Cell Viability Assay to perform Cell cytotoxicity / Proliferation assay cell type - HT-22

Get tips on using Porcine Endothelial Cell Growth Medium to perform Mammalian cell culture media PCAEC

Get tips on using Porcine Endothelial Cell Growth Medium to perform Mammalian cell culture media PAOEC

Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCAEC

Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCtASMC

Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCtAEC

Get tips on using Endothelial Cell Growth Medium MV to perform Mammalian cell culture media BAOEC

Get tips on using Cell-APOPercentage™ Apoptosis Assay to perform Apoptosis assay cell type - SKOV3