siRNA / miRNA gene silencing Human HT-29

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary human aortic smooth muscle cells

Get tips on using miRNeasy Serum/Plasma Kit to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

Transfection is a powerful technique that enables the study of the function of genes and gene products in cells. Based on the nature of experiments, we may need a stable DNA transfection in cells for persistent gain-of-function or loss-of-function of the target gene. For stable transfection, integration of a DNA vector into the chromosome is crucial which requires selective screening and clonal isolation. By carefully selecting a viral delivery system and related reagents we can ensure safe and highly-efficient delivery of expression constructs for high-level constitutive or inducible expression in any mammalian cell type.

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 Nitrocef disks to perform Reporter gene assay β-lactamase substrates - HEK 293 & HEK 293T cells

Get tips on using FGF-10 Antibody (3C7): sc-293208 to perform Immunohistochemistry Human - FGF-10

Get tips on using EpiScope MSP Kit to perform DNA methylation profiling Gene specific profiling - HEK 293T NELL2

Get tips on using EpiScope MSP Kit to perform DNA methylation profiling Gene specific profiling - HEK 293T NELL1

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - HEK 293T RHO

Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Gene specific profiling - HEK 293T RBP3