Labeled Tubulin-Alexa Fluor® 647
Catalog Number: 064705
Molecular Weight: ~110 kDa
Maximum Excitation/Emission: 651/672 nm
Labeling Stoichiometry: ~0.5 (check product label)
Form: blue aqueous solution
Concentration: 20 mg/ml
Buffer Conditions: 50 mM K-Glutamate and 0.5 mM MgCl2 (pH 7.0)
Shipping: shipped on dry ice
Storage Conditions: store at -80˚C immediately
Shelf Life: check product label for expiration date
Taxol Stabilized Microtubules (long and flexible)
Taxol-stabilized microtubules at 5 mg/ml total tubulin protein with Labeled Tubulin-Alexa Fluor® 647 (Cat. No. 064705) and Cycled Tubulin™ (Cat. No. 032005) at a 1:5 ratio. Images acquired on a widefield fluorescence microscope. View protocol.
Labeled Tubulin-Alexa Fluor® 647 is generated by reacting Alexa Fluor® 647 succinimidyl ester with >99% pure tubulin protein, thereby covalently linking the dye to random tubulin surface lysines. Polymerization competency is maintained during the labeling process by reacting the tubulin protein with the dye in its polymerized form and subjecting it to a final polymerization/depolymerization cycle. The final labeled tubulin protein displays maximum absorption at 280 nm and 651 nm, and a labeling stoichiometry ([dye]/[tubulin]) of ~0.5 as determined by spectroscopic analysis (Figure 1). Specific labeling stoichiometries are indicated on the product label. Labeled Tubulin-Alexa Fluor® 647 is commonly visualized with a Cy5 filter set with maximum excitation/emission wavelengths of 651/672 nm (see images above). 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® 647 will polymerize into microtubules when supplemented with guanosine-5'-triphosphate (GTP), warmed to 37˚C, and kept above its critical concentration. Polymerization activity is detectable in a variety of experimental systems including fluorescence microscopy assays, turbidity assays, and GTPase assays. Labeled tubulin protein is suitable for use in a variety of fluorescent experimental applications including live cell injection, and can be combined with unlabeled tubulin protein in generating fluorescent microtubules in vitro. Cycled Tubulin™ (Cat. No. 032005) is highly recommended as the unlabeled tubulin protein source in order to minimize tubulin heterogeneity, as labeled tubulin reagents are derived directly from Cycled Tubulin™. 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.
The Brightest Far Red Tubulin Protein On The Market
Labeled Tubulin-Alexa Fluor® 647 displays maximum absorption at 280 nm and 651 nm. Spectroscopic analysis reveals a peak at 280 nm and 651 nm, indicating absorption by the tubulin protein and Alexa Fluor® 647 dye, respectively.