Cell cytotoxicity / Proliferation assay

Get tips on using anti-p62 Protein, C-Terminal Specific Polyclonal Antibody to perform Autophagy assay cell type - MDA-MB-231

Get tips on using eBioscience™ Annexin V-FITC Apoptosis Detection Kit to perform Apoptosis assay cell type - SMMC-7721, HEPG2

Get tips on using FITC Annexin V Apoptosis Detection Kit I (RUO) to perform Apoptosis assay cell type - SMMC-7721, HEPG2

Get tips on using eBioscience™ Annexin V-FITC Apoptosis Detection Kit to perform Apoptosis assay cell type - Caspase 3/7

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - MLO-Y4

Get tips on using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb #3398 to perform Autophagy assay cell type - HEK 293

Get tips on using anti-p62 / SQSTM1 (C-terminus) guinea pig polyclonal, serum to perform Autophagy assay cell type - CaCo-2

Get tips on using FITC Annexin V Apoptosis Detection Kit with 7-AAD to perform Apoptosis assay cell type - PANC-1

Get tips on using Cell DNA Isolation Kit to perform DNA isolation / purification Cells - Immortalized cell lines C2C12

Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) have been greatly used for studies on embryonic development and cell differentiation.iPSCs provide a stable source for either self-renewal or differentiation into suitable cells when cultured in a particular environment. Pluripotent cell culture was originally started by deriving cells from inner cell mass (ICM) from pre-implanted blastocysts, these were called embryonic stem cells. These cells after isolation can be grown on traditional extracellular matrices (like mouse embryonic fibroblasts, MEFs) or feeder-free culture systems. DMEM/F12 has been the most commonly used basal media in the culture of pluripotent cells. These cells are cultured at normal atmospheric oxygen levels, 21%, however, some studies have proposed that 4% oxygen tension may be better for hESC growth. Higher D-glucose concentration (4.2g/l) and osmolarity (320mOsm) that mimics the natural environment of embryonic tissue are optimal for the growth of hESCs. Supplements like N2 and/or B-27, in the presence of growth factors like bFGF, have been shown to increase pluripotency of these cells. bFGF, FGF2 and other ligands of receptor tyrosine kinases like IGF are also required or maintain self-renewal ability of these cells. TGF𝛃1, by its activation of SMAD2/3 signalling, also represses differentiation of iPSCs. Other compounds like ROCK inhibitors reduce blebbing and apoptosis in these cells to maintain their clonogenicity. However, an inhibitor for LIF (leukaemia inhibitory factor, which is one of the pluripotent genes) has an opposing effect. Therefore, it is important to understand the culture conditions and media composition that affect downstream signalling in hESCs or iPSCs that may lead to their differentiation.