Minute™ Plant Golgi Apparatus Enrichment Kit
The plant Golgi apparatus plays an important role in biosynthesis of cellular structural components and
protein trafficking. However proteomic characterization of the Golgi has been hindered by limited
methodologies for obtaining enriched/purified Golgi fraction. Traditional method for isolating Golgi apparatus
depends upon the use of density centrifugation, which is tedious and time consuming. Large amount of
starting material is usually required. Plant Golgi stacksare notoriously labile and loss of stack integrity can
occur if harsh homogenization is used. This means that assessment of purity by morphological means such
as electron microscopy is difficult. This kit is designed to overcome certain disadvantages of traditional
methods by employing spin column-based homogenization, differential centrifugation and selective
precipitation for enrichment of the Golgi. The protocol is simple and straight forward and requires only
milligram amount of starting material. Significantenrichment of Golgi fraction (2-3 folds) can be obtained.
Enrichment of Golgi Apparatus from Plant Leaves
A. Ponceau S stained blot. B. Western blot with ER (BIP2), Golgi (Arf1) and histone 3 (H3) marker antibodies.
The antibodies were purchased from Agrisera (Vannas, Sweden). Lanes 1 & 2: total protein from A. thaliana
and N. tabacum; Lanes 3 & 5: enriched Golgi fractions from A. thaliana and N. tabacum, respectively.