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Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis
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نویسنده
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fontaine a. ,pascual a. ,diouf i. ,bakkali n. ,bourdon s. ,fusai t. ,rogier c. ,almeras l.
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منبع
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parasites and vectors - 2011 - دوره : 4 - شماره : 1
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چکیده
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Mosquito salivary proteins are involved in several biological processes that facilitate their blood feeding and have also been reported to elicit an igg response in vertebrates. a growing number of studies have focused on this immunological response for its potential use as a biological marker of exposure to arthropod bites. as mosquito saliva collection is extremely laborious and inefficient,most research groups prefer to work on mosquito salivary glands (sgs). thus,sg protein integrity is a critical factor in obtaining meaningful data from immunological and biochemical analysis. current methodologies rely on an immediate freezing of sgs after their collection. however,the maintenance of samples in a frozen environment can be hard to achieve in field conditions. in this study,sg proteins from two mosquito species (aedes aegypti and anopheles gambiae s.s.) stored in different media for 5 days at either +4°c or room temperature (rt) were evaluated at the quantitative (i.e.,elisa) and qualitative (i.e.,sds-page and immunoblotting) levels. our results indicated that pbs medium supplemented with an anti-protease cocktail seems to be the best buffer to preserve sg antigens for 5 days at +4°c for elisa analysis. conversely,cell-lysis buffer (urea-thiourea-chaps-tris) was best at preventing protein degradation both at +4°c and rt for further qualitative analysis. these convenient storage methods provide an alternative to freezing and are expected to be applicable to other biological samples collected in the field. © 2011 fontaine et al; licensee biomed central ltd.
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آدرس
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unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France, unite de recherche en biologie et epidemiologie parasitaires - umr6236 - ifr48,antenne marseille de l'institut de recherche biomédicale des armées (irba),le pharo,bp 60109,13 262 marseille cedex 07, France
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Authors
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