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文献和实验fractions between 48 and 60 % (w/w) sucrose. 5 ul of fraction + 5 ml PE - mix well and pellet onto coverslips: 12,500 rpm for 15' at 4 deg.C in HB-4. Post fix in methanol (-20 deg.C) for 5' and rehydrate and do 5051 + antitubulin followed by anti-mouse, anti
tubes (cold) Solutions: A lot of these solutions are w/w; Tim says that to make these weigh out sucrose and then add buffer till weight is 100g. Wash & Lysis: PE: 10 mM PIPES, 1mM EDTA, 8 mM BME Make a 50X stock
Intracellular Cytokine Staining Protocol
)2d/3danti-CD3 (10ug/ml immobilized)+anti-CD28 (2ug/ml soluble) 5hrMonensinJES6-5H4IL-4mouse spleenConA (3ug/ml) (2d)/IL-2 (20ng/ml)+IL-4 (20ng/ml) (3d)2d/3danti-CD3 (10ug/ml immobilized)+anti-CD28 (2ug/ml soluble) 5hrMonensinBVD6-24G2IL-6mouse
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