siRNA / RNAi /miRNA transfection Human Cells Cal 27 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.

Get tips on using QIAamp DNA Mini Kit to perform DNA isolation / purification Cells - Primary cells Cyst-derived kidney epithelial cells

Get tips on using QIAamp DNA FFPE Tissue Kit to perform DNA isolation / purification Cells - Primary cells Pseudomyxoma peritonei (PMP) cells

Get tips on using Human ShhN ELISA to perform ELISA Human - ShhN

Get tips on using Human Melanocyte Media to perform Mammalian cell culture media HEM

Get tips on using IntestiCult™ Organoid Growth Medium (Human) to perform 3D Cell Culture Media Human cancer colon organoids

Get tips on using ApopTag® Peroxidase In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - HNSCC Detroit 562 human head and neck tumor cells

Get tips on using Human ANGPTL3 ELISA to perform ELISA Human - Angiopoietin-Like 3 (AngptL3)

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.