dna-isolation-purification-cells-primary-cells-mouse-embryonic-fibroblast-mef

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Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Bacteria - Gram positive Enterococcus faecium

Products Qiagen DNeasy Blood & Tissue Kit

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Bacteria - Gram positive Bacillus subtilis

Products Qiagen DNeasy Blood & Tissue Kit

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Bacteria - Gram positive Weissella confusa

Products Qiagen DNeasy Blood & Tissue Kit

Get tips on using DNeasy PowerLyzer Microbial Kit (50) to perform DNA isolation / purification Bacteria - Gram negative Rhodopseudomonas

Products Qiagen DNeasy PowerLyzer Microbial Kit (50)

Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Immortalized cell lines MDA-MB-231

Products Polyplus transfections jetPEI® DNA transfection, HTS application

Stem cells have the unique ability to self-renew or differentiate themselves into various cell types in response to appropriate signals. These cells are especially important for tissue repair, regeneration, replacement, or in the case of hematopoietic stem cells (HSCs) to differentiate into various myeloid populations. Appropriate signals refer to the growth factor supplements or cytokines that mediate differentiation of various stem cells into the required differentiated form. For instance, HSCs can be differentiated into dendritic cells (with IL-4 and GM-CSF), macrophages (with m-CSF) and MDSCs (with IL-6 and GM-CSF). Human pluripotent stem cells (hPSCs) and induced pluripotent stem cells (iPSCs) can be first cultured in neural differentiation media (GSK3𝛃-i, TGF𝛃-i, AMPK-i, hLIF) to form neural rosettes, which can be differentiated into neural or glial progenitors (finally differentiated into oligodendrocytes). Neural progenitors can be finally differentiated into glutaminergic (dibytyryl cAMP, ascorbic acid) and dopaminergic (SHH, FGF-8, BDNF, GDNF, TGF-𝛃3) neurons. Thus, it is important to first identify the self-renewing cell line: its source and its final differentiation state, followed by the supplements and cytokines required for the differentiation, and final use. Timelines are another thing that is considered. For instance, it takes 7-10 days to form neural rosettes from iPSCs and 3 days to differentiate neural progenitors to neurons. Finally, the stability for stem cell culture media varies. It is advised to make fresh media every time when differentiating HSCs to myeloid populations, whereas neural differentiation media may remain stable for two weeks when stored in dark between 2-8C.

Cell culture media Stem cell Differentiation media hiPSCs differentiation into mesodermal lineage cells

Get tips on using Gibco™Neurobasal™ Medium to perform Stem cell culture media NCH421K cells primary glioma

Products Thermo Fisher Scientific Gibco™Neurobasal™ Medium

Get tips on using Senescence Cells Histochemical Staining Kit to perform Reporter gene assay β-galactosidase substrates - adipose stem cells

Products Sigma-Aldrich Senescence Cells Histochemical Staining Kit

Get tips on using MethylMiner Mehtylated DNA Enrichment Kit to perform DNA methylation profiling Whole genome profiling - mouse iPSCs

Products Thermo Fisher Scientific MethylMiner Mehtylated DNA Enrichment Kit

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

Cellular assays Autophagy assay cell type K562 cells

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