Comparison of three methods of protein extraction from Dermatophagoides pteronyssinus for two-dimensional electrophoresis / 中国医学科学杂志(英文版)
Chinese Medical Sciences Journal
;
(4): 64-68, 2009.
Article
in English
| WPRIM
| ID: wpr-302647
ABSTRACT
<p><b>OBJECTIVE</b>To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis (2-DE) analysis.</p><p><b>METHODS</b>The extracts of Dermatophagoides pteronyssinus were prepared with Coca's solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid (BCA) assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis.</p><p><b>RESULTS</b>The concentrations of extracted protein by methods of Coca's solution, lysis buffer, and Trizol reagent were 0.63 g/L, 0.90 g/L, and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight (LMW) range could be detected with the Coca's solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight (MMW) protein spots were absent. Several MMW protein spots (174-178 kD and 133 kD) and more LMW protein spots were detected with Trizol reagent method.</p><p><b>CONCLUSIONS</b>Among Coca's solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Phenols
/
Allergens
/
Electrophoresis, Gel, Two-Dimensional
/
Proteins
/
Chemistry
/
Dermatophagoides pteronyssinus
/
Guanidines
Limits:
Animals
Language:
English
Journal:
Chinese Medical Sciences Journal
Year:
2009
Type:
Article
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