Direkt zum InhaltDirekt zur SucheDirekt zur Navigation
▼ Zielgruppen ▼

IRI Life Sciences

Humboldt-Universität zu Berlin | IRI Life Sciences | News | New resources online: How to purify active, label-free tubulin and how to visualize microtubule dynamics in vitro

New resources online: How to purify active, label-free tubulin and how to visualize microtubule dynamics in vitro

The Reber lab published two protocols on how to purify active, label-free tubulin and on how to visualize microtubule dynamics in vitro in STAR protocols. Both are #openaccess

The group of Simone Reber at IRI Life Sciences has been invited to publish two articles in STAR Protocols, which are connected to their publication in Current Biology earlier this year. Original publication: Hirst WG, Biswas A, Mahalingan KK and Reber S. (2020) “Differences in intrinsic tubulin dynamic properties contribute to spindle length control in Xenopus species”.

Affinity-Purification of Label-free Tubulins from Xenopus Egg Extracts
Sebastian Reusch, Abin Biswas, William Graham Hirst, and Simone Reber
DOI: 10.1016/j.xpro.2020.100151

Cytoplasmic extracts from unfertilised Xenopus eggs have made important contributions to our understanding of microtubule dynamics, spindle assembly, and scaling. Until recently, these in vitro studies relied on the use of heterologous tubulin. This protocol details the purification of tubulin from egg extracts of three different Xenopus species, which yields milligrams of physiologically relevant tubulins. The purified tubulin is a complex mixture of isoforms with various post-translational modifications specific to the source, i. e. eggs of different frog species. The protocol is applicable to any cell or tissue of interest.

Graphical Abstract: Tubulin purification

Graphical Abstract: Tubulin purification (© Reusch S, Biswas A, Hirst WG, Reber S)


In vitro Reconstitution and Imaging of Microtubule Dynamics by Fluorescence and Label-free Microscopy
William Graham Hirst, Christine Kiefer, Mohammad Kazem Abdosamadi, Erik Schäffer, Simone Reber
DOI: 10.1016/j.xpro.2020.100177

Imaging microtubule dynamics can be instructive for a given species, isoform composition or biochemical modification. Here, we describe step by step two methods that visualize microtubule dynamics at high speed and high contrast: (1) Total internal reflection fluorescence microscopy (TIRFM) and (2) label-free interference reflection microscopy (IRM).

Graphical Abstract: Total internal reflection fluorescence microscopy

Graphical Abstract: Total internal reflection fluorescence microscopy (© Hirst WG, Kiefer C, Abdosamadi MK, Schäffer E, Reber S)


Contact
Further information and requests for resources and reagents should be directed to Simone Reber via email: simone.reber@iri-lifesciences.de.