Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform DNA isolation / purification Bacteria - Gram positive Lactobacillus
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 Purified Rat Anti-Mouse CD31 to perform Immunohistochemistry Mouse - CD31
Get tips on using Biotin Rat Anti-Mouse CD24 to perform Immunohistochemistry Mouse - CD24
Get tips on using Mouse/Rat Neuropilin-1 Antibody to perform Immunohistochemistry Mouse - Nrp1
Get tips on using Mouse RANKL ELISA Kit (TNFSF11) (ab100749) to perform ELISA Mouse - RANK L
Get tips on using Wizard® Plus Maxipreps DNA Purification System to perform Plasmid Isolation E. coli DH5α
Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase
Get tips on using KAPA RNA HyperPrep Kit with RiboErase (HMR) to perform RNA sequencing Mouse - ESCs (Embryonic Stem Cells)
Get tips on using Anti-Mouse CD31 (PECAM-1) to perform Immunohistochemistry CD31 - Rat Mouse -NA-
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