Figure 1. Isolation of γ-TuCs bound to... | Rockefeller University Press

$Figure 1. Isolation of γ-TuCs bound to γ-TuNA. (A) Schematic outline of the isolation procedure. (B) After gradient centrifugation, an aliquot of each fraction was resolved by SDS-PAGE followed by silver staining. Proteins resolved from the peak fraction of γ-TuRC (Fr. 10) were identified by mass spectrometry. The contaminant protein above GCP6 also appeared in the precipitates of blank beads. (C) The gradient fractions were analyzed by immunoblotting. (D) In a replicate gel stained with Sypro ruby, the relative amounts of γ-tubulin and GCPs were determined from the isolated γ-TuRC (Fr. 10) to derive the stoichiometry. The ratios of proteins to GCP5 are presented as mean ± SD from three independent experiments.$

Figure 1.

Isolation of γ-TuCs bound to γ-TuNA. (A) Schematic outline of the isolation procedure. (B) After gradient centrifugation, an aliquot of each fraction was resolved by SDS-PAGE followed by silver staining. Proteins resolved from the peak fraction of γ-TuRC (Fr. 10) were identified by mass spectrometry. The contaminant protein above GCP6 also appeared in the precipitates of blank beads. (C) The gradient fractions were analyzed by immunoblotting. (D) In a replicate gel stained with Sypro ruby, the relative amounts of γ-tubulin and GCPs were determined from the isolated γ-TuRC (Fr. 10) to derive the stoichiometry. The ratios of proteins to GCP5 are presented as mean ± SD from three independent experiments.

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