Protein expression and purification Tissue

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Tissue - mouse uterus tissue

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Tissue - mouse ovary tissue

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Tissue - mouse oviduct tissue

Get tips on using Magna ChIP™ G Tissue Kit to perform ChIP Human - MDA-MB-231

Get tips on using DNA Isolation Kit for Cells and Tissues to perform DNA isolation / purification Cells - Primary cells Human primary keratinocytes

Get tips on using PureLink™ RNA Mini Kit to perform RNA isolation / purification Tissue - mouse adipose tissue

Get tips on using pET30a(+)-karp to perform Protein Expression Prokaryotic cells - E. coli 56‐kDa O. tsutsugamushi strain Karp protein

Get tips on using pTip-QC2-gi_21221796 to perform Protein Expression Prokaryotic cells - R. erythropolis putative tetR-family transcriptional regulatory protein

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.

Get tips on using pcDNA™3.1 (+) Mammalian Expression Vector to perform siRNA / miRNA gene silencing Human - U251 cofilin-1 (CFL1)