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文献和实验Labeling Tubulin with Tetramethylrhodamine Succinimidyl Ester
to ~37℃ by soaking in warm water.If possible,keep a separate centrifuge and SS34 rotor at 4℃. 5.PEM buffer: 0.1 M PIPES,1.0 mM EGTA,0.5 mM MgCl2,pH 6.9,prepare 50 ml.Keep ~20 ml at 37℃ and the rest on ice. 6.DMSO. 7.TRSE,Molecular Probes C-1171.Prepare
LABELING MUSCLE ACTIN WITH CARBOXYFLUORESCEIN SUCCINIMIDYL ESTER
. 4.3 mM CaCl2,1.2 M KI,50 mM Tris-HCl,pH 8.0,5 ml. 5.0.5 mM ATP,0.2 mM CaCl2,0.5 mM DTT,2 mM Tris-HCl,pH 8.0 at 4℃,1 liter. 6.CFSE (Molecular Probes,C-1311; not the diacetate),prepare a stock solution of 50 mg/ml in DMSO. 7.Small SS34 tubes and adaptors
Phycoerythrin conjugation protocol
concentrator (Centricon 30 or 100 are fine) Apply to top of separation column. Elute with PBS. Pump column at approximately 0.3 ml/min. Order of elution is conjugate first (MW ~400,000) then free PE (MW ~240,000) then free Ab (MW ~150,000
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