biotin labeled tubulin  (Cytoskeleton Inc)

Journal: Nature Communications

Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau

doi: 10.1038/s41467-025-67888-x

Figure Lengend Snippet: A Snapshots showing the dynamic gliding of microtubules ( cyan , labeled with HiLyte 488,) along DNA ( green , stained with SYTOX Orange), driven by the mobility of tau condensates ( magenta , labeled with Cy5) on DNA. Microtubules were formed by incubating with 5 μM tubulin. B Schematic illustration of the DNA–tau–microtubule tripartite assembly. C Representative kymograph displaying the mobility of the gliding microtubule on DNA shown in ( A ). D Time-lapse images capturing the pivoting motion of the microtubule, anchored by tau–DNA co-condensates. In ( A ) and ( B ), red boxes mark the location of the DNA strands, and white dashed lines show the position of the microtubules in the preceding image. E – G TIRF images of high-density surface-tethered λ-DNA ( green , stained with SYTOX Orange) before tau addition ( E ), after adding 5 μM tau ( magenta , Cy5-labeled) with 5% PEG ( F ), and after adding a mixture of tau and tubulin ( cyan , HiLyte 488-labeled) with 5% PEG ( G ). Microtubules were pulled to the surface by tau–DNA co-condensates. H , I Large-area views of microtubule pulldown on DNA surfaces with and without preformed tau–DNA co-condensates. In ( H ), microtubules did not attach to the DNA surface without tau pre-incubation to form condensates; only background fluorescence from out-of-focus tubulins was visible. J Quantification of surface-bound microtubules with and without preformed tau–DNA co-condensates ( n = 9 images; mean ± s.d.).

Article Snippet: After co-condensate formation, HiLyte 488-labeled tubulin (Cytoskeleton, #TL488M-A) was mixed with unlabeled tubulin (Cytoskeleton, #T240-B), making up 4% of the total tubulin concentration.

Techniques: Labeling, Staining, Incubation, Fluorescence