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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2017.6(12): 2968-2975
DOI: https://doi.org/10.20546/ijcmas.2017.612.346


A Simple, Rapid and Effective Protocol for Extraction of Total Plant Proteins from Cotton Leaf
Anirudha Kumar Sahu1, I.S. Katageri1*, M.P. Jadhav1 and H.M. Vamadevaiah2
1Department of Biotechnology
2Department of Biochemistry, College of Agriculture, University of Agricultural Sciences, Dharwad-580005, Karnataka, India
*Corresponding author
Abstract:

An attempt was made to conceive a simple, rapid and effective method to extract total plant proteins from cotton leaf samples for proteomic analysis; particularly SDS PAGE. The protocol combines TCA/acetone precipitation, methanol washes and phenol extraction as reported by Wang et al., (2006) with few modifications to separate the proteins from several interfering compounds like lipids, polysaccharides, phenolics, terpenes, pigments, organic acids, nucleic acids, free ions, proteolytic and oxidative enzymes; and precipitate high-quality protein from a very low amount of starting material. The key components of this protocol were water soluble PVP and DTT (those prevent oxidation of phenolic compounds); and PMSF which acted as a protease inhibitor and the key step in this protocol was the phenol extraction step. The overall quality of the protein was good with less vertical streaking and smearing in the SDS PAGE gel and the average protein yield obtained was approximately 510 g/g fresh weight of cotton leaf.


Keywords: TCA/acetone precipitation, Methanol washes, Phenol extraction, SDS PAGE, Cotton leaf.
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How to cite this article:

Anirudha Kumar Sahu, I.S. Katageri, M.P. Jadhav and Vamadevaiah, H.M. 2017. A Simple, Rapid and Effective Protocol for Extraction of Total Plant Proteins from Cotton Leaf.Int.J.Curr.Microbiol.App.Sci. 6(12): 2968-2975. doi: https://doi.org/10.20546/ijcmas.2017.612.346