Alexa Fluor 594 - GMPCCP Stabilized (short, rigid microtubules); Taxol Stabilized (long, flexible microtubules)

Figure 1: Labeled Tubulin-Alexa Fluor® 594 is commonly visualized with a Texas Red filter set. Left, GMPCPP-stabilized microtubules at 2 mg/ml [tubulin] with Labeled Tubulin-Alexa Fluor® 594 (Cat. No. 059405) and Cycled Tubulin™ (Cat. No. 032005) at a 1:10 ratio. Right, taxol-stabilized microtubules at 5 mg/ml [tubulin] with Labeled Tubulin-Alexa Fluor® 594 (Cat. No. 059405) and Cycled Tubulin™ (Cat. No. 032005) at a 1:10 ratio. Images acquired on a widefield fluorescence microscope.


alexa-594-tubulin

Figure 2: Labeled Tubulin-Alexa Fluor® 594 displays maximum absorption at 280 nm and 590 nm. Spectroscopic analysis reveals a peak at 280 nm and 590 nm, indicating absorption by tubulin and Alexa Fluor® 594, respectively.

LABELED TUBULIN-ALEXA FLUOR® 594

Catalog Number: 059405
Source: Bovine Brain

Material
Labeled Tubulin-Alexa Fluor® 594 is commonly visualized with a Texas Red filter set with maximum excitation/emission wavelengths of 590/617 nm (Figure 1). The labeled tubulin is generated by reacting Alexa Fluor® 594 succinimidyl ester (Thermo Fisher A20104) with Cycled Tubulin™ (Cat. No. 032005), thereby covalently linking the dye to random tubulin surface lysines. Cycled Tubulin™ is >99% pure and polymerization competent. These properties are maintained during the labeling process by reacting Cycled Tubulin™ with the dye in its polymerized form and subjecting it to a final polymerization/depolymerization cycle. The final product displays maximum absorption at 280 nm and 590 nm, and a labeling stoichiometry ([dye]/[tubulin]) of 0.5-2.0 as determined by spectroscopic analysis (Figure 2). Specific labeling stoichiometries are indicated on the product label. The product is cryopreserved at 20 mg/ml in 50 mM K-Glutamate and 0.5 mM MgCl2 (pH=7.0).

Activity and Applications
Labeled Tubulin-Alexa Fluor® 594 will polymerize into microtubules when supplemented with GTP, warmed to 37˚C, and kept above its critical concentration. Labeled tubulin is suitable for use in a variety of fluorescent experimental applications including live cell injection, and can be combined with unlabeled tubulin (Cycled Tubulin | Cat. No. 032005) in generating fluorescent microtubules in vitro. Visit our protocols page for common microtubule polymerization protocols, including the generation of short, rigid microtubules stabilized by GMPCPP or long, flexible microtubules stabilized by taxol.

  • in vitro microtubule gliding assays
  • single molecule kinesin and dynein motor assays
  • speckle microscopy
  • live cell injection
  • in vitro nanoscale devices
  • TIRF, STORM, SIM, STED, TPE, confocal, and widefield microscopy

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