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A high sensitivity ...
A high sensitivity dot-blot assay for proteoglycans by cuprolinic blue precipitation.
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- Jortikka, Matti (författare)
- epartment of Anatomy, University of Kuopio, Kuopio, Finland
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- Lammi, Mikko, 1961- (författare)
- epartment of Anatomy, University of Kuopio, Kuopio, Finland,Chondrogenic and Osteogenic Differentiation Group
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- Parkkinen, Jyrki (författare)
- epartment of Anatomy, University of Kuopio, Kuopio, Finland
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- Lahtinen, Risto (författare)
- epartment of Anatomy, University of Kuopio, Kuopio, Finland
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- Tammi, Markku (författare)
- epartment of Anatomy, University of Kuopio, Kuopio, Finland
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(creator_code:org_t)
- Informa Healthcare, 1993
- 1993
- Engelska.
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Ingår i: Connective Tissue Research. - : Informa Healthcare. - 0300-8207 .- 1607-8438. ; 29:4, s. 263-272
- Relaterad länk:
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http://www.tandfonli...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- A highly sensitive blot-assay was developed for glycosaminoglycans (GAGs) and proteoglycans (PGs) utilizing a precipitation reaction by a cationic dye Cuprolinic Blue. The precipitates were deposited into 1-2 mm2 spots on nitrocellulose membrane by using a 96-well filtration apparatus. The dried sheet was digitized by a flat bed scanner and the intensity of the dots was quantitated by an image analysis software. The working range for chondroitin sulfate was 10-300 ng. The response of various GAGs differed according to the number of anionic groups, both sulphate and carboxyl groups being able to bind the dye. The sensitivity of the assay was decreased by high concentrations of GuC, CsC and protein, but not by nonionic detergents, common buffers and 8 M urea. Contact exposure to autoradiography film enabled quantitation of 25-250 DPM, and 1-10 DPM, of 35SO4-radioactivity in precipitated PGs after overnight and 14 days' exposures, respectively.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- Proteoglycans
- glycosaminoglycans
- dot-blot assay
- Cuprolinic acid
- image analysis
- analytisk kemi
- Analytical Chemistry
- biokemi
- Biochemistry
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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