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. 1997 Dec 1;139(5):1271–1280. doi: 10.1083/jcb.139.5.1271

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Measurement of the dissociation constant (K_d) for full length Stu2p. A constant amount of ³⁵S-labeled in vitro translated Stu2p (∼10⁻¹¹ M final concentration) was incubated with variable amounts of taxol-stabilized bovine brain microtubules. Microtubules were pelleted by centrifugation, and both bound Stu2p (in pellets) and unbound Stu2p (in supernatants) were subjected to SDS-PAGE analysis. Band intensities were quantitated using the phosphoimager software. (A) Autoradiographs of Stu2p in the supernatants (S) and in the corresponding pellets (P). Microtubule concentration ranging from 0 to 5 μM is shown above each lane. (B) The binding curve was generated from the above data. The K_d is equal to the concentration of polymerized tubulin required to cosediment 50% of the Stu2p in the reaction.