ImageJ=1.50g

Cycled Tubulin™ (Cat. No. 032005) at 1.5 mg/ml with Alexa-488 Tubulin at 1:10 growing off of GMPCPP stabilized microtubule seeds (red).

CYCLED TUBULIN™ (>99%)

Catalog Number: 032005
Source: Bovine Brain

Material

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).

Activity

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.

Uses

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

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Figure 1: Cycled Tubulin™ is >99% pure. Coomassie G250-stained protein gel of Cycled Tubulin™ separated by SDS-PAGE. The tubulin appears as a single species migrating at ~55 kDa. Molecular weight markers (kDa) and loaded protein quantities are indicated.


Cycled Tubulin Purity PurSolutions

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.

Cycled Tubulin Polymerization Turbidity Freeze-Thaw PurSolutions

Compare with Lyophilized Tubulin