Western blot p62/SQSTM1 Mouse IgG2b Human

Get tips on using QIAamp DNA Blood Mini Kit to perform DNA isolation / purification Bacteria - Gram positive Staphylococcus aureus

Get tips on using Gentra Puregene Blood Kit to perform DNA isolation / purification Cells - Primary cells Bone marrow mononuclear cells

Get tips on using Gentra Puregene Blood Kit to perform DNA isolation / purification Cells - Immortalized cell lines Lymphoblastoid cell lines

Get tips on using QIAamp DNA Blood Mini Kit to perform DNA isolation / purification Cells - Immortalized cell lines HEK 293T

Get tips on using DNeasy Blood and Tissue Kit (250) to perform DNA isolation / purification Cells - Immortalized cell lines Loucy

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Bacteria - Gram positive piezophilic bacteria [AT7 and AT12 Strains]

Get tips on using FitAmp Blood and Cultured Cell DNA Extraction Kit to perform DNA isolation / purification Cells - Immortalized cell lines SH-SY5Y

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Though DNA quantification is but one small step in the multifaceted DNA sample preparation workflow, it can have large implications on the performance and validity of conclusions drawn from downstream assays. Major challenges include accuracy, precision, reproducibility, and detection of present contamination. Among UV spectrophotometry, fluorescence and real-time PCR based methods, the quantification method should be chosen based on the requirement of the downstream assay.

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.