siRNA / miRNA gene silencing Human Melanoma cells (501 Mel and SK Mel 28)

Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10

Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10

Get tips on using pSilencer™ 4.1-CMV neo to perform shRNA gene silencing Human - SiHa AEG-1

Get tips on using GATA-1 shRNA Plasmids (h) to perform shRNA gene silencing Human - TF‐1 GATA‐1

Get tips on using HiPerFect Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - H9c2 Cationic and neutral lipids

Get tips on using TurboFect Transfection Reagents to perform siRNA / RNAi /miRNA transfection Rat - H9c2 Cationic and neutral lipids

Get tips on using RNAqueous®-Micro Total RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized SK-BR-3

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

Acid phosphatase detection heavily relies on determining the concentration of tartrate-resistant acid phosphatase (TRAP) in the sample. Hence, sample preparation is very crucial and it should be done strictly as per kit manufacturer instructions to avoid any inconsistency and poor sensitivity