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© 1996 Springer-Verlag. Reproduced with permission.
(from Durso et al. 1996, Protoplasma 190: 141-150)
Dual immunolocalizations using
conventional epifluorescence microscopy on footprints of untreated
(a & b), APM-treated (c & d),
and taxol-treated (e & f) carrot protoplasts
lysed prior to fixation are shown. Fluorescein fluorescence reporting
affinity-purified anti-pp50 antibodies (a, c, & e)
is highly similar to Texas Red fluorescence reporting anti-carrot
tubulin mAb1F8 (b, d, & f). Controls using the
two fluorochrome-conjugated secondary antibodies alone or together
produced no visible fluorescence (not shown). Moreover, single
localizations using either of the primary antibodies in combinations
with either of the secondary antibodies demonstrated that the
two excitation and emission wavelengths are exclusively specific
for their respective detection filter sets (not shown). These
data indicate that pp50 co-localizes with microtubules in interphase
cortical arrays, and that the co-localization is microtubule-dependent.
Bars = 4 um.
