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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.