Published: Vol 3, Iss 10, May 20, 2013 DOI: 10.21769/BioProtoc.569 Views: 8644
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Abstract
Mannans are hemicellulosic polysaccharides and are present in cell walls of all land plants. Mannan polysaccharides are synthesized by two enzymes, mannan synthase (ManS) for backbone (mannan or glucomannan) synthesis and galactomannan galactosyl transferase for side-chain (galactosyl) addition. Here, a method for ManS activity assay using microsomes freshly isolated from Arabidopsis stems is described. This method can be applied to isolation of microsomes from any tissues of Arabidopsis or any other plants.
Keywords: ArabidopsisMaterials and Reagents
Equipment
Procedure
4x ManS assay buffer | 10 μl | (final concentration in reaction: 1x) |
1 mM GDP-Man | 0.85 μl | (final concentration in reaction: 21.18 μM) |
GDP-[14C]-Man(76.34 μM) | 2 μl | (final concentration in reaction: 3.82 μM) |
Deionized H2O | 7.15 μl |
Recipes
Acknowledgments
This protocol is adapted from Wang et al. (2013).
References
Article Information
Copyright
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Wang, Y. (2013). Stem Microsome Preparation and Mannan Synthase Activity Assay. Bio-protocol 3(10): e569. DOI: 10.21769/BioProtoc.569.
Category
Biochemistry > Carbohydrate > Polysaccharide
Biochemistry > Protein > Activity
Cell Biology > Organelle isolation > Microsome
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