Get tips on using Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System to perform Reporter gene assay β-galactosidase substrates - U87 and U251 glioblastoma cells
Get tips on using QuikChange Multi Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Human - Point mutation BxPC-3 uPAR
Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - PC-3 human prostate cancer
Get tips on using ApopTag Fluorescein in Situ Apoptosis Detection Kit to perform TUNEL assay cell type - PC-3 human prostate cancer
The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.
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.
Get tips on using BrdU Cell Proliferation Assay Kit to perform Cell cytotoxicity / Proliferation assay cell type - BxPc-3 human primary pancreatic adenocarcinoma
Get tips on using Low Input Quick Amp Labeling Kits to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Cyanine 3-pCp
Get tips on using Ovation® RNA Amplification System V2 to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Cyanine 3-pCp
Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - PANC-, BxPC-3 human pancreas
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