siRNA / miRNA gene silencing Human Melanoma cells (501 Mel and SK Mel 28)

Get tips on using Gibco™ MEM α, GlutaMAX™ Supplement, no nucleosides to perform Stem cell Differentiation media Human oogonial stem cells differentiation into oocytes

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.

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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 Cell Staining Kit (BioVision) to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells

Get tips on using LIVE/DEAD™ Cell Imaging Kit to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells

Get tips on using Neural Progenitor Medium 2 to perform Stem cell Differentiation media Differentiation of Human PSC into Neural progenitor cells

Get tips on using Dulbecco’s Modified Eagle’s Medium - high glucose to perform Stem cell culture media Human Dental pulp stem cells (hDPSC)

Get tips on using Gibco™Essential 8™ Medium to perform Stem cell Differentiation media hiPSC differentiation into Human Neuronal cells

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Human lung fibroblasts