Chapter title |
Depletion of RuBisCO Protein Using the Protamine Sulfate Precipitation Method
|
---|---|
Chapter number | 17 |
Book title |
Proteomic Profiling
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2550-6_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2549-0, 978-1-4939-2550-6
|
Authors |
Ravi Gupta, Sun Tae Kim, Gupta, Ravi, Kim, Sun Tae |
Abstract |
Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is a major high-abundant protein (HAP) in the plant leaves which hinders analysis of low-abundant proteins (LAP). In this chapter, we describe a highly simple RuBisCO depletion method using protamine sulfate (PS). Addition of 0.1 % PS is sufficient to precipitate the RuBisCO from the leaf extracts of diverse plants including monocots and dicots. Our results of SDS-PAGE, Western blotting, and two-dimensional gel electrophoresis showed that both large and small subunits of RuBisCO were precipitated in the pellet fractions, while LAPs were enriched in the supernatant fraction after PS precipitation. |
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