Cat # PG-049
Manual & Protocol | MSDS
The plant Golgi apparatus plays an important role in the 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. The traditional method for isolating Golgi apparatus depends upon the use of density centrifugation, which is tedious and time-consuming. A large amount of starting material is usually required. Plant Golgi stacks are 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 the enrichment of the Golgi. The protocol is simple and straightforward and requires only a milligram amount of starting material. Significant enrichment of the 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&4: enriched Golgi fractions from A. thaliana and N. tabacum, respectively.
Kit includes:
Buffer A
10 ml
Buffer B
1.0 ml
Buffer C
4 ml
Buffer D
0.5
Filter Cartridge with Collection Tubes
20 units
Plastic Rods
2
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