Evaluation of 2-D DIGE for skeletal muscle: Protocol and repeatability

Malm, Christer (author): Umeå universitet,Institutionen för integrativ medicinsk biologi (IMB),Department of Intergrative Medical Biology, Section for Anatomy, Umeå University, Sweden; Winternet, Boden, Sweden

Hadrevi, Jenny (author): Högskolan i Gävle,Umeå universitet,Idrottsmedicin,Centrum för belastningsskadeforskning,Umeå University, Department of Integrative Medical Biology, Section for Anatomy, Umeå, Sweden

Bergström, Sven-Anders (author): Umeå universitet,Medicin,Department of Public Health and Clinical Medicine, Umeå University, Sweden

Pedrosa-Domellof, Fatima (author): Umeå universitet,Institutionen för integrativ medicinsk biologi (IMB),Oftalmiatrik,Department of Integrative Medical Biology, Section for Anatomy, Umeå University

Antti, Henrik (author): Umeå universitet,Kemiska institutionen,Department of Chemistry, Section for Organic Chemistry, Umeå University, Sweden

Svensson, Michael (author): Umeå universitet,Idrottsmedicin,Department of Surgical and Preoperative Sciences, Section for Sports Medicine, Umeå University, Sweden

Frängsmyr, Lars (author): Umeå universitet,Institutionen för integrativ medicinsk biologi (IMB),Department of Intergrative Medical Biology, Section for Anatomy, Umeå University, Sweden and Department of Surgical and Preoperative Sciences, Section for Sports Medicine, Umeå University, Sweden

(creator_code:org_t)

2009-07-08
2008
English.
In: The Scandinavian Journal of Clinical & Laboratory Investigation. - : Informa Healthcare. - 0036-5513 .- 1502-7686. ; 68:8, s. 793-800

Related links:: https://urn.kb.se/re...; show more...; https://doi.org/10.1...; https://urn.kb.se/re...; show less...

Abstract Subject headings

Proteomic analysis has the potential to yield vast amounts of data. The available proteomic methods have been hampered by methodological errors in quantification due to large gel-to-gel variations. The inclusion of an internal standard greatly reduces this variation, and therefore the purpose of this investigation was: 1) to develop a sample preparation protocol for human skeletal muscle for two-dimensional differentiated gel electrophoresis (DIGE) and 2) to investigate the repeatability of one particular system, the Ettan™ DIGE. To test repeatability, nine aliquots from the same homogenate were labelled with three different CyDye™ dyes (Cy2, Cy3, Cy5). Samples were run on 1824 cm gels, scanned with a Typhoon™ 9410 laser scanner and analysed in the DeCyder™ software. When selecting spots appearing only in triplicate (n = 1314), the mean error was 1.7 % (SD: 10.5 %; 95 % CI: 1.1-2.4 %). When setting the significance level to 99 %, no false-positive changes in protein volume ratios were detected. In the protocol presented here, only 0.5 mg tissue was used and separation of >2500 distinct protein spots in the pH range 3-11 and MW 10-200 kDa. Changes in protein abundance of <20 % could be detected. The method is especially useful when comparing muscle proteins between different conditions; for example, healthy and diseased tissue, before and after treatment or different exercise protocols.

MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Oftalmologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Ophthalmology (hsv//eng)
MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Clinical Medicine (hsv//eng)
MEDICIN OCH HÄLSOVETENSKAP -- Hälsovetenskap (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Health Sciences (hsv//eng)

By the author/editor: Malm, Christer; Hadrevi, Jenny; Bergström, Sven- ...; Pedrosa-Domellof ...; Antti, Henrik; Svensson, Michae ...; show more...; Frängsmyr, Lars; show less...

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Clinical Medicin ...; and Ophthalmology

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Basic Medicine; and Cell and Molecul ...

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