Immunohistochemistry Collagen VII antibody [LH7.2] Mouse Human

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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 GeneChip® Human Genome U133 Plus 2.0 Array to perform RNA amplification & labeling Mammalian - RNA amplification and Labeling Human Endometrial Stromal cells Biotin

Get tips on using GeneChip® Human Genome U133 Plus 2.0 Array to perform RNA amplification & labeling Mammalian - RNA, rhesus monkey brain tissue Human endothelail stromal cells

Get tips on using SiRNA silencing human Eph receptor B4, Id: s243 to perform siRNA / miRNA gene silencing Human - HNSCC cell line Eph receptor B4 Polymer / Lipid

Get tips on using SiRNA silencing human Eph receptor B4, Id: 533 to perform siRNA / miRNA gene silencing Human - HNSCC cell line Eph receptor B4 Polymer / Lipid

Get tips on using SurePrint G3 Human Gene Expression 8x60K v2 Microarray Kit to perform Microarray Human - PCOS

Get tips on using Human Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit to perform ELISA Human - ShhN

Get tips on using Human Retinol binding protein 4 ELISA Kit (RBP4) (ab108897) to perform ELISA Human - RBP4

Get tips on using Human NRG1-beta 1/HRG1-beta 1 DuoSet ELISA to perform ELISA Human - NRG1