摘要:Considerations for useThe bicinchoninic acid (BCA)assay is available in kit form from Pierce (Rockford,Ill.).This procedure is very applicable to microtiter plate methods.The BCA is used for the same [阅读全文:]
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摘要:Considerations for useThe principle of the biuret assay is similar to that of the Lowry,however it involves a single incubation of 20 min.There are very few interfering agents (ammonium salts being on[阅读全文:]
摘要:Considerations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay.It is fairly accurate and samples that are out of range can be retested within minute[阅读全文:]
摘要:Materials1.Drop frozen 2x cycled tubulin in a polymerization-promoting buffer.Stored at -80℃2.GTP,100 mM stock,titrate to pH ~7.Need 2 ml.3.Waterbath,set at 37℃4.SS34 rotor,5 ml and 15 ml tubes and ad[阅读全文:]
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摘要:Materials1.2 mM PIPES,pH 7.0,500 ml.2.Gc-globulin (Calbiochem 345802),1 mg.3.CFSE (Carboxyfluorescein succinimidyl ester; Molecular Probes).4.Lysine,100 mM in buffer 1,10 ml.5.G25-150 desalting column[阅读全文:]
摘要:Materials1.Small vial and stir bar.2.Bio-Beads SM-2 in 0.7x15 cm column.3.2 mM Tris-HCl,pH 8.5,4℃,250 ml.4.Centricon-30 concentrator.5.5 mM Tris-acetate,0.1 mM DTT,pH 6.95,4℃,250 ml.6.200 mM K-borate,[阅读全文:]
摘要:Materials1.200 mM K-borate,pH 9.0,10 ml.2.100 mM lysine,100 mM K-borate,pH 9.0,1 ml aliquots stored at -20℃.3.2 mM Tris-HCl,pH 8.5,4℃,250 ml.4.100 mM DTT stock.5.Centricon-30 concentrator.6.5 mM Tris-[阅读全文:]
摘要:Day 0 and 1Materials1.0.5 mM ATP,0.2 mM CaCl2,2 mM Tris-HCl,pH 8.0 at 4℃,250 ml for day 0.2.0.5 mM ATP,2 mM MgCl2,100 mM boric acid,pH 8.3 at 4℃,10 ml.3.N-(1-pyrene)iodoacetamide (Molecular probes P-2[阅读全文:]