Non-Cycled Tubulin Drug Binding PurSolutions

Lyophilized Tubulin (Cat. No. 142001) is ideal for use in turbidity assays and high-throughput screening applications.

LYOPHILIZED TUBULIN (>99%)

Catalog Number: 142001
Source: Bovine Brain

Material

Lyophilized Tubulin was isolated by selective precipitation from bovine brain homogenate by an adaptation of the method of Andreu (2007) and lyophilized by an adaptation of the method of Dráberová et al. (2010). The resulting product is >99% pure (Figure 1) and polymerization competent (Figure 2). Lyophilized Tubulin is supplied as a white powder with a tubulin-to-powder weight ratio of 1:3. When reconstituted with ultrapure water to 40 mg/ml, the buffer conditions are 10 mM Sodium Phosphate, 0.5 mM MgCl2, 0.1 mM GTP, and 0.25 M Trehalose, pH 7.0.

Activity

When supplemented with guanosine (GTP or GMPCPP) and warmed to 37°C, Lyophilized Tubulin will polymerize into MTs when above its critical concentration. The recommended tubulin concentration for ensuring polymerization is 2 mg/ml.

Uses

Lyophilized 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: Lyophilized Tubulin is >99% pure. Coomassie G250-stained protein gel of Lyophilized 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.


Non-Cycled Tubulin Purity PurSolutions

Figure 2: Lyophilized Tubulin is polymerization-competent. Absorbance (340 nm) of Lyophilized 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 at 37°C. Distinct nucleation and polymerization phases are evident.

lyophilized-tubulin-polymerization

Compare with Cycled Tubulin™