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Sample extraction techniques for enhanced proteomic analysis of plant tissues
Year:
2006
Source of publication :
Nature Protocols
Authors :
Isaacson, Tal
;
.
Volume :
1
Co-Authors:
Isaacson, T., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Damasceno, C.M.B., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Saravanan, R.S., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
He, Y., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Catalá, C., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Saladié, M., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Rose, J.K.C., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Facilitators :
From page:
769
To page:
774
(
Total pages:
6
)
Abstract:
Major improvements in proteomic techniques in recent years have led to an increase in their application in all biological fields, including plant sciences. For all proteomic approaches, protein extraction and sample preparation are of utmost importance for optimal results; however, extraction of proteins from plant tissues represents a great challenge. Plant tissues usually contain relatively low amounts of proteins and high concentrations of proteases and compounds that potentially can limit tissue disintegration and interfere with subsequent protein separation and identification. An effective protein extraction protocol must also be adaptable to the great variation in the sets of secondary metabolites and potentially contaminating compounds that occurs between tissues (e.g., leaves, roots, fruit, seeds and stems) and between species. Here we present two basic protein extraction protocols that have successfully been used with diverse plant tissues, including recalcitrant tissues. The first method is based on phenol extraction coupled with ammonium acetate precipitation, and the second is based on trichloroacetic acid (TCA) precipitation. Both extraction protocols can be completed within 2 d.
Note:
Related Files :
article
chemistry
isolation and purification
methodology
plant
Plant Proteins
Plants
Proteome
proteomics
vegetable protein
Show More
Related Content
More details
DOI :
10.1038/nprot.2006.102
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
20724
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:38
Scientific Publication
Sample extraction techniques for enhanced proteomic analysis of plant tissues
1
Isaacson, T., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Damasceno, C.M.B., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Saravanan, R.S., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
He, Y., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Catalá, C., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Saladié, M., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Rose, J.K.C., Department of Plant Biology, Cornell University, 228 Plant Sciences Building, Ithaca, NY 14853, United States
Sample extraction techniques for enhanced proteomic analysis of plant tissues
Major improvements in proteomic techniques in recent years have led to an increase in their application in all biological fields, including plant sciences. For all proteomic approaches, protein extraction and sample preparation are of utmost importance for optimal results; however, extraction of proteins from plant tissues represents a great challenge. Plant tissues usually contain relatively low amounts of proteins and high concentrations of proteases and compounds that potentially can limit tissue disintegration and interfere with subsequent protein separation and identification. An effective protein extraction protocol must also be adaptable to the great variation in the sets of secondary metabolites and potentially contaminating compounds that occurs between tissues (e.g., leaves, roots, fruit, seeds and stems) and between species. Here we present two basic protein extraction protocols that have successfully been used with diverse plant tissues, including recalcitrant tissues. The first method is based on phenol extraction coupled with ammonium acetate precipitation, and the second is based on trichloroacetic acid (TCA) precipitation. Both extraction protocols can be completed within 2 d.
Scientific Publication
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