siRNA / miRNA gene silencing Human Jurkat MK2 (MAPK Kinase 2)

Get tips on using X-tremeGENE™ siRNA Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - AR42J Lipid based

Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Mouse - 3T3-L1 S6 kinase 1

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 CD31-FITC, 5.6E, 2 mL, ASR to perform Flow cytometry Anti-bodies Human - CD31/PECAM-1

Get tips on using TACS® 2 TdT Fluorescein Kit to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using EMBacY#2 to perform Protein Expression Eukaryotic cells - Hi5 hcAbl

Get tips on using pDDGFP-2 to perform Protein Expression Eukaryotic cells - S. cerevisiae GPCRs

Get tips on using TACS® 2 TdT Fluorescein Kit to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Monkey - Point mutation Vero UL23 thymidine kinase

Get tips on using Lipofectamine® RNAiMAX Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - OV-2008 Lipofectamine