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Identification of the major proteins of an immune modulating fraction from adult Fasciola hepatica released by Nonidet P40.

Veterinary parasitology (2012-10-02)
Russell M Morphew, Clare M Hamilton, Hazel A Wright, David J Dowling, Sandra M O'Neill, Peter M Brophy
RESUMEN

Fasciola hepatica NP-40 released protein extract (FhNPE) exhibits potent Th1 immunosuppressive properties in vitro and in vivo. However, the protein composition of this active fraction, responsible for Th1 immune modulatory activity, has yet to be resolved. Therefore, FhNPE, a Nonidet P-40 extract, was subjected to a proteomic analysis in order to identify individual protein components. This was performed using an in house F. hepatica EST database following 2D electrophoresis combined with de novo sequencing based mass spectrometry. The identified proteins, a mixture of excretory/secretory and membrane-associated proteins, are associated with stress response and chaperoning, energy metabolism and cytoskeletal components. The immune modulatory properties of these identified protein(s) are discussed and HSP70 from F. hepatica is highlighted as a potential host immune modulator for future study.

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Sigma-Aldrich
IGEPAL® CA-630, for molecular biology
Supelco
IGEPAL® CA-630
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IGEPAL® CA-630, viscous liquid
Sigma-Aldrich
Triton X-114, laboratory grade
Sigma-Aldrich
Triton X-114
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Triton X-405 solution, 70% in H2O
Sigma-Aldrich
Triton X-165 solution, Non-ionic, CMC-0.43 mM, suitable for electrophoresis, cloud point >100 °C
Sigma-Aldrich
IGEPAL® CA-720, average Mn ~735