gmpcpp Protocols

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Recycling Tubulin Protocol - http://mitchison.med.harvard.edu/protocols/recycle.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Recycle tubulin fractions stored at -80¡C after the PC column and store the recycled tubulin in small aliquots for day-to-day use. Generally store recycled tubulin in Injection Buffer (IB) without free GTP. This is done because depolymerization appears to be much better in IB, IB is ideal for microinjections/adding tubulin to extracts, and the absence of free GTP makes polymerization with GMPCPP, a very useful GTP analog that has ~5-10X lower affinity than GTP for tubulin. - [Read Recycling Tubulin Protocol]

Tubulin Polymerization with GTP/GMPCPP/Taxol Protocol - http://mitchison.med.harvard.edu/protocols/poly.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Protocol for tubulin polymerization with GTP/GMPCPP/ Taxol. Includes: Solutions & Supplies; Prepolymerization Clarification; GTP Polymerization; Taxol Polymerization; GMPCPP Polymerization; Determining Concentration of GMPCPP/Taxol MTs. - [Read Tubulin Polymerization with GTP/GMPCPP/Taxol Protocol]

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Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.