• Fluorescent-based readout for tubulin polymerization-360 nm (ex) / 420 nm (em)
• Cost-effective HTS screen for anti-cancer compounds targeting microtubule dynamics
• Characterization of microtubule-binding proteins (MAPs)
• IC50 & EC50 determinations for anti-tubulin ligands

Kit contents (sufficient for 96 assays)

• Tubulin protein (>99% pure)
• Tubulin buffer with fluorescent reporter
• Tubulin glycerol buffer
• Guanosine triphosphate (GTP)
• Paclitaxel (polymerization enhancer)
• DMSO
• Half area 96-well, black, flat bottom microtiter plate
• Detailed protocol

Equipment & materials required

• Protein or compound of interest
• Temperature-controlled fluorimeter with 360 ±20 nm (ex) / 420 ± 20 nm (em)

This assay measures tubulin polymerization dynamics using a fluorescent reporter that incorporates into microtubules as they form. It is designed to monitor the real-time kinetics of microtubule assembly and disassembly, which is critical for understanding cellular functions and screening pharmacological agents.

Key features

• Fluorescent-based readout - 360 nm (ex) / 420 nm (em)
• 100 µg tubulin per assay/15% glycerol, 96 well plates, 96 assays
• Can be adapted to 384 well plates, 20 µg per assay, 480 assays
• 50 µl volume per assay, 96-well format
• 10 µl volume per assay , 384-well format
• Coefficient of variation (CV): 11%
• Paclitaxel EC50: 0.49 µM under standard conditions
• Vinblastine IC50: 0.6 µM under standard conditions