Site Directed Mutagenesis (SDM) Human Deletion PC-3

Get tips on using ON-TARGETplus human ATG16L1 siRNA to perform siRNA / miRNA gene silencing Human - SHSY5Y ATG16L1

Get tips on using MISSION® esiRNA_ human CCL2 to perform siRNA / miRNA gene silencing Human - U251 CCL2

Get tips on using ON-TARGETplus Human TET1 siRNA to perform siRNA / miRNA gene silencing Human - A172 TET1

Get tips on using Silencer® FANCD2 siRNA (human) to perform siRNA / miRNA gene silencing Human - 501 Mel and SK Mel 28 FANCD2

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Human breast cancer MCF-7 cells-Mammospheres

Get tips on using ON-TARGETplus Human ARL2BP siRNA to perform siRNA / miRNA gene silencing Human - HeLa BART/ARL2BP

Get tips on using ON-TARGETplus Human BECN1 siRNA to perform siRNA / miRNA gene silencing Human - U251 Beclin 1

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres

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.