CYCLED TUBULIN™ (>99%)
Catalog Number: 032005
Source: Bovine Brain
Cycled Tubulin™ is isolated by cycling bovine brain homogenate through conditions that promote tubulin polymerization/depolymerization in high salt buffers by an adaptation of the method of Castoldi and Popov (2003). The resulting product is >99% pure (Figure 1) and polymerization competent (Figure 2). Cycled Tubulin™ is cryopreserved at 20 mg/ml in Tubulin PEM Buffer (Cat. No. 032002; 80 mM PIPES, 1 mM EGTA, and 1 mM MgCl2, pH 6.8).
When supplemented with guanosine (GTP or GMPCPP) and warmed to 37°C, Cycled Tubulin™ will polymerize into MTs when above its critical concentration. The recommended tubulin concentration for ensuring polymerization is 2 mg/ml.
Cycled Tubulin™ is supplied for use in cell-free experimental systems including:
- structural analysis by X-ray crystallography and electron microscopy
- drug discovery by high-throughput screening
- in vitro biochemical and biophysical approaches
Greater purity at a better price.
Figure 2: Cycled Tubulin™ is polymerization-competent. Absorbance (340 nm) of Cycled Tubulin™ at 5 mg/ml in Tubulin PEM Buffer (Cat. No. 032002; 80 mM PIPES, 1 mM EGTA, and 1 mM MgCl2, pH 6.8) supplemented with 1 mM GTP, 1 mM DTT, and 5% Glycerol at 37°C. Distinct nucleation and polymerization phases are evident. Purple, product that has undergone one freeze-thaw cycle. Blue, product that has undergone two freeze-thaw cycles.