Immunohistochemistry Anti-Glial Fibrillary Acidic Protein (GFAP) Mouse Human

Get tips on using FlowCellect Autophagy LC3 antibody based kit to perform Autophagy assay cell type - K562 cells

Get tips on using PCNA Antibody (F-2) to perform Cell cytotoxicity / Proliferation assay cell type - oral squamous cell carcinoma

Get tips on using OxPhos Complex IV subunit IV Monoclonal Antibody (20E8C12) to perform Western blotting COX4

Get tips on using Phospho-SMAD2 (Ser465, Ser467) Monoclonal Antibody (H.205.4) to perform Western blotting Smad2

Get tips on using Oct-3/4 Antibody (C-10): sc-5279 to perform Western blotting Oct4

Get tips on using ERK 1/2 Antibody (C-9): sc-514302 to perform Western blotting ERK

Get tips on using β-2-Microglobulin Antibody (BBM.1): sc-13565 to perform Western blotting β₂ microglobulin

Get tips on using p-c-Jun Antibody (KM-1): sc-822 to perform Western blotting C-Jun

Get tips on using PI 3-kinase p85α Antibody (B-9) to perform Autophagy assay cell type - SH-SY5Y

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.