siRNA / miRNA gene silencing Human Primary Human Aortic Endothelial Cells GLO-1

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray Human - Precision cut lung slices Target preparation kit (RNA Amplification + Hybridization + control)

Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - SKOV3, Caov3 human ovarian cancer

Get tips on using ApoBrdU Red DNA Fragmentation Kit to perform TUNEL assay cell type - SKOV3, Caov3 human ovarian cancer

Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - SK-MEL-2 human melanoma

Get tips on using H2A.X Phosphorylation Assay Kit (Flow Cytometry) to perform DNA Damage Assay Human Skin Fibroblast Cell (FSK)

Get tips on using CytoSelect™ 24-Well Wound Healing Assay to perform Wound healing assay cell type - human HMEC

Get tips on using CytoSelect™ 24-Well Wound Healing Assay to perform Wound healing assay cell type - human HUVEC

Get tips on using EZ-Magna ChIP™ A/G Chromatin Immunoprecipitation Kit to perform ChIP Human - Fibroblast 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.