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文献和实验Proteasome Preparation from Human Brain
10% 1ml 9ml Gently load 7ml of each % into MSE tube for 6x38 swing out rotor (highest % first) 4) On top of gradient, load 5ml supernatant. 5) Balance tubes and spin at 23,000 rpm
CaPO3 Transfection in 293 T cells
before harvesting. Check pH of buffers (critical). Combine 100X NaPi with 2X HEPES in a 1:50 ratio, so that the final concentration of NaPi and HEPES is 2X. Filter sterilize all buffers in the cell culture hood.
Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC
coverslips with acetone for 15'-30', followed by ethanol for 15', and then spin drying them works well for VE-DIC. Other labs use far more extensive and excruciating cleaning procedures. For fluorescence assays coverslips can be used straight from a box
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