dc.contributor.author | Grant, Ryan | |
dc.contributor.author | Inal, Jameel | |
dc.date.accessioned | 2018-07-31T13:02:47Z | |
dc.date.available | 2018-07-31T13:02:47Z | |
dc.date.issued | 2012-09-13 | |
dc.identifier.citation | Grant , R & Inal , J 2012 , ' Isolation of microvesicles and exosomes by microfiltration and estimation of normal reference range in blood plasma. ' , Microvesiculation and Disease, a Biochemical Society Focused Meeting , London , United Kingdom , 13/09/12 - 14/09/12 . | |
dc.identifier.citation | conference | |
dc.identifier.other | PURE: 13382473 | |
dc.identifier.other | PURE UUID: 6b4ffe32-0aa1-4d40-a726-37f6bb64797d | |
dc.identifier.uri | http://hdl.handle.net/2299/20313 | |
dc.description.abstract | Current protocols for the isolation of microvesicles (MVs) and exosomes, which in the main focus on differential centrifugation, vary considerably (1). In an attempt to set a new standard, we describe a filtration protocol for isolating phosphatidylserine-positive MVs (larger than 100 nm in diameter) and exosomes. The key preparative step to successfully isolate both MVs and exosomes to a high degree of purity was a gentle sonication to break up exosome clumps. Filtration through a 100 nm pore size Millipore filter allowed for collection of exosomes in the filtrate. The larger MVs could then be recovered from the filter. Annexin V-PE MVs were sized and quantified using Polysciences Polybead Microspheres (200 nm) and BDTrucount tubes, respectively on a FACS CaliburTM flow cytometer. The normal reference range from normal human donors was found to be 0.51-2.82 x105 MVs/ml. Freeze/thawing of samples had little effect on MV counts and with age MV levels seemed only marginally reduced. Fasting status also affected MV levels, appearing up to 3-fold higher in fasting individuals. Smokers had lower MV counts and nicotine reduced MV release from THP-1 cells. | en |
dc.language.iso | eng | |
dc.title | Isolation of microvesicles and exosomes by microfiltration and estimation of normal reference range in blood plasma. | en |
dc.contributor.institution | School of Life and Medical Sciences | |
dc.contributor.institution | Department of Biological and Environmental Sciences | |
dc.contributor.institution | Biosciences Research Group | |
dc.description.status | Peer reviewed | |
rioxxterms.version | AM | |
rioxxterms.type | Other | |
herts.preservation.rarelyaccessed | true | |