• In vitro microtubule pelleting assay for identification of MAPs
• Detection of MAPs using recombinant proteins or cell extracts
• Biochemical characterization of MAPs

Kit content (50 - 100 assays)

• Tubulin protein (99% pure)
• MAP fraction protein
• Bovine serum albumin
• General tubulin buffer
• Cushion buffer
• Salt extraction buffer
• GTP
• Paclitaxel
• DMSO

Equipment & materials required

• Reagents for detection of target protein (e.g., Coomassie or antibody)
• Ultracentrifuge: 100,000g, 4°C & 37°C, 50-200 µl volume tube capacity

This assay takes advantage of the fact that microtubules (MTs) pellet when centrifuged at 100,000 × g. Any protein bound to MTs will co-pellet during centrifugation. When using purified proteins, comparing the supernatant and pellet fractions via SDS-PAGE is sufficient to determine MT association. When working with cell lysates or in vitro translation products, detection can be performed using western blotting.

Key characteristics

• Assay type: Primarily qualitative (Yes/No); quantitative analysis possible through titration assays
• Detection method: SDS-PAGE or Western blot
• Timeframe: Approximately 4–6 hours