CFSE kit来自于thermo, 以下为实验步骤: CFSE assay For new CFSE tube, add 18 ul DMSO into the tube, to get 5 mM (working solution 1:1,000 dilute) 1 mL complete RPMI for 10e6 cell; 0.5 ml for < 5e6 cell 1.1 ul 5mM in 110 ul PBS In tube bottom, put 1ml cells In tube top, ...
接着，执行T细胞活化步骤，使用thermo的CD3/CD28 dynabeads激活T细胞。最终结果如图所示，红色曲线代表对照组，绿色部分表示活化的T细胞。每次增殖后，CFSE荧光强度会减少一半。这就是CFSE T细胞增殖实验的全过程，是不是非常简便呢？
TheCFSEshouldbestableforatleast12monthsifstoreat-20°C,protectingfromlightandmoisture, avoidingfreeze/thawcycles. ChemicalandPhysicalProperties MolecularWeight:557.46 Solvent:Dimethylsulfoxide(DMSO) SpectralProperties:Ex/Em=494/521nm AssayProtocol Note:Followingisourrecommendedprotocolforlivecells.Itonlyprovidesa...
Protocol: Using CellTrace Violet with T cells Protocol: Positive magnet isolation of CD4+ T cells from human PBMCs Poster: A Comparison of Three Techniques to Induce Efficient Ex Vivo T Cell Expansion Usage...
Unlike other techniques that measure a static parameter of a specific time-point, CFSE staining allows to distinguish between subsequent cell divisions. Here, we show a simple protocol to study human T and NK cell proliferation with CFSE dilution assay by flow cytometry....
cell cycle analysis, apoptosis assays such as annexin V staining, CellEvent Caspase-3/7 assay, and TUNEL assay, cell viability, proliferation assays such as CellTrace assay and Click-iT EdU assay, measurements of mitochondrial potential with MitoProbe assays, and cell counting using counting beads...
A CFSE based assay for measuring CD4+CD25+ regu- latory T cell mediated suppression of auto-antigen specific and polyclonal T cell responses. J. Immunol. Methods 322: 1-11.Venken, K., Thewissen, M., Hellings, N., Somers, V., Hensen, K., Rum- mens, J. L. and Stinissen, P., ...
Conclusions Phenytoin-specific proliferation may be detected with greater sensitivity by the CFSE dilution assay than the conventional LTT. The assay revealed that both CD4+ and CD4 T cells proliferated and produced IFN-γ and TNF-α after stimulation with phenytoin. The CFSE dilution assay might ...
Human PBMCs, Human T Cells Primary Incubation 5µM, stock diluted in PBS, 20 Mins @ 37°C, protected from light Blocking Agent None Secondary Incubation for assessment of T-cell proliferation: 5 days, assay-dependent Tertiary Incubation ...
6 ASSAY PROTOCOL ASSAY PROTOCOL 7 CFSE Labeling of Cells 1. Centrifuge the cells at 300 x g for five minutes at room temperature and aspirate the supernatant. 2. Wash cells with pre-warmed sterile PBS and centrifuge as above. 3. Resuspend cell pellet in pre-warmed PBS at a concentration ...