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In vitro EB1 microtubule binding. Cell extracts from J77 prepared in buffer A (see Materials and Methods) were incubated with taxol-stabilized microtubules. Supernatants (S) and pellets (P) were collected and separated by SDS-polyacrylamide gel, transferred to poly(vinylidene difluoride), and probed with the anti-EB1 antibody GD10. Microtubule binding was demonstrated by cosedimentation of EB1 with the microtubule pellet. Whole-cell lysate (L) demonstrated endogenous EB1 mobility.
