general tubulin buffer  (Cytoskeleton Inc)

Journal: Nature Communications

Article Title: Decoupling phase separation and fibrillization preserves activity of biomolecular condensates

doi: 10.1038/s41467-026-69244-z

Figure Lengend Snippet: a Schematic of tubulin recruitment and MT polymerization in Tau condensates. b Nascent SynTag-Tau condensates (age = 2 h) in the presence of 50 nM HiLyte647-labeled tubulin show enrichment of tubulin in the dense phase. Corresponding line profiles are shown below. c Aged SynTag-Tau condensates (age = 4 h) with the addition of 50 nM HiLyte647-labeled tubulin show altered tubulin partitioning to the condensate interface. Corresponding line profiles are shown below. Display ranges in images of (b) and (c) were adjusted independently for better clarity. d Condensate age-dependent MT polymerization assay in SynTag-Tau condensates. e MT surface coverage plot corresponding to panel (d). Horizontal lines at each time point represent the median value. The individual data points represent measurements based on three independent replicates. Sample size: 1 hr, 30 datapoints; 2.5 hr, 24 datapoints; 5 hr, 23 datapoints; 8 hr, 24 datapoints; 12 hr, 16 datapoints; 16 hr, 14 datapoints. f Frequency-domain (FD) FLIM map of SynTag-Tau condensates at various time points. g Representative fluorescence lifetime distributions at 1 h and 5 h time points (since sample preparation). The points are the data, and the lines are Gaussian fittings with the corresponding goodness of fit (R 2 ) indicated. h FD-FLIM map of aged SynTag-Tau condensates with emergent amyloid fibrils (sample age = 8 h). i Schematic of condensate physical aging-induced interfacial resistance in SynTag-Tau condensates, and its effect on tubulin partitioning and MT assembly. Each of these experiments was independently repeated at least three times. Source data are provided as a Source Data file.

Article Snippet: Briefly, SynTag-Tau protein solution was buffer-exchanged to general tubulin buffer (Cytoskeleton) consisting of 80 mM PIPES (pH 6.9), 2 mM MgCl 2 , and 0.5 mM EGTA, supplemented with 2 mM DTT.

Techniques: Labeling, Polymerization Assay, Fluorescence, Sample Prep

Journal: Nature Communications

Article Title: Decoupling phase separation and fibrillization preserves activity of biomolecular condensates

doi: 10.1038/s41467-026-69244-z

Figure Lengend Snippet: a MT polymerization assay of SynTag-Tau condensates treated with 1 mM L-Arg at various time points. b A comparison of MT surface coverage as a function of condensate age between untreated SynTag-Tau condensates (Fig. ) and L-Arg treated SynTag-Tau condensates (images corresponding to 2 mM condition are shown in Supplementary Fig. ). Horizontal black lines represent the median value. The individual data points represent measurements based on three independent replicates. Sample size for untreated condition: 1h, 30 datapoints; 2.5h, 24 datapoints; 5h, 23 datapoints; 8 h, 24 datapoints; 12 h, 16 datapoints; 16 h, 14 datapoints. For 1 mM L-Arg condition: 1h, 19 datapoints; 2.5h, 15 datapoints; 5 h, 16 datapoints; 8 h, 15 datapoints; 12 h, 11 datapoints; 16 h, 13 datapoints. For 2 mM L-Arg condition: 1 h, 20 datapoints; 2.5 h, 19 datapoints; 5 h, 15 datapoints; 8 h, 11 datapoints; 12 h, 11 datapoints; 16 h, 15 datapoints. Statistical significance was determined using an unpaired two-sided Student’s t -test between the MT surface coverages of untreated condensates (salmon) and either 1 mM L-Arg treated condensates (green) or 2 mM L-Arg treated condensates (blue) (* means p < 0.05, ** means p < 0.01, ***means p < 0.001, ****means p < 0.0001). The associated P values are shown from left to right: 0.0028, 0.0003, 0.0264, 0.0006, 0.0002, and 0.00009. c Schematic representing the proposed model of biochemical modulation of condensate metastability governing protein condensate-to-fibril conversion and its impact on the condensate biochemical activity in tubulin partitioning (shown as dimers) and MT formation (shown as tubule). These experiments were independently repeated three times. Source data are provided as a Source Data file.

Article Snippet: Briefly, SynTag-Tau protein solution was buffer-exchanged to general tubulin buffer (Cytoskeleton) consisting of 80 mM PIPES (pH 6.9), 2 mM MgCl 2 , and 0.5 mM EGTA, supplemented with 2 mM DTT.

Techniques: Polymerization Assay, Comparison, Activity Assay