Microtiter plate assay for biopterin using cryopreserved Crithidia fasciculata
This chapter presents microtiter plate assay for biopterin, using cryopreserved Crithidia fasciculate. The term “total biopterin” includes biopterin, dihydrobiopterin, and tetrahydrobiopterin. Most of the diagnostic criteria and normative data available for the Crithidia assay have been obtained by...
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| Veröffentlicht in: | Methods in Enzymology 1997, Vol.281, p.61-70 |
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| creator | Leeming, Robert J. |
| description | This chapter presents microtiter plate assay for biopterin, using cryopreserved Crithidia fasciculate. The term “total biopterin” includes biopterin, dihydrobiopterin, and tetrahydrobiopterin. Most of the diagnostic criteria and normative data available for the Crithidia assay have been obtained by automated tube assay without chloramphenicol. This requires heating to remove protein, and although heat treatment is not necessary for the microtiter plate assay, the practice has continued for comparative, diagnostic purposes. Plasma samples are diluted 1:30 and whole-blood samples are diluted 1:40 in phosphate buffer. Dried blood spots have an 8-ram circle punched out into 1 ml of phosphate buffer. Samples are then heated in a steamer for 10 min, centrifuged, and passed through 0.2-μm filters to clarify. When there are reasons to suspect that the biopterin level is raised, for example, in phenylketonuria, further dilutions should be prepared in phosphate buffer after filtration. |
| doi_str_mv | 10.1016/S0076-6879(97)81009-9 |
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The term “total biopterin” includes biopterin, dihydrobiopterin, and tetrahydrobiopterin. Most of the diagnostic criteria and normative data available for the Crithidia assay have been obtained by automated tube assay without chloramphenicol. This requires heating to remove protein, and although heat treatment is not necessary for the microtiter plate assay, the practice has continued for comparative, diagnostic purposes. Plasma samples are diluted 1:30 and whole-blood samples are diluted 1:40 in phosphate buffer. Dried blood spots have an 8-ram circle punched out into 1 ml of phosphate buffer. Samples are then heated in a steamer for 10 min, centrifuged, and passed through 0.2-μm filters to clarify. 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When there are reasons to suspect that the biopterin level is raised, for example, in phenylketonuria, further dilutions should be prepared in phosphate buffer after filtration.</description><subject>Animals</subject><subject>Biological Assay - methods</subject><subject>Biopterins - analogs & derivatives</subject><subject>Biopterins - analysis</subject><subject>Biopterins - biosynthesis</subject><subject>Biopterins - blood</subject><subject>Crithidia fasciculata - growth & development</subject><subject>Cryoprotective Agents</subject><subject>Humans</subject><subject>Molecular Structure</subject><subject>Phenylalanine - blood</subject><subject>Phenylketonurias - metabolism</subject><subject>Reference Values</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><issn>0076-6879</issn><issn>1557-7988</issn><isbn>9780121821821</isbn><isbn>012182182X</isbn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9UdtOwzAMjbhoTGOfMKlPCB4KSdrUyRNCEzdpwAPwHGWpA0FdW5J20v6e7iIsS5Z8ji37HEJmjF4zyoqbd0qhSAsJ6lLBlWSUqlQdkTETAlJQUh6TqQJJGWdylydk_D9yRqYx_tAhBEBOsxEZKS6oKmBMXl-8DU3nOwxJW5kOExOj2SSuCcnSN-3Q93XSR19_JTZsmjZgxLDGMpkH33370pvEmWi97Ydpc05OnakiTg91Qj4f7j_mT-ni7fF5frdIkRe8S3PugGeZkrktnQMnc7SlUcYaAdw5IVhGM5FLsCUoruQSjETHsYCCM3RZNiEX-71taH57jJ1e-WixqkyNTR81KCZFQeVAnB2I_XKFpW6DX5mw0QcBBvx2j-Nw7dpj0MMvWFssfUDb6bLxmlG9NUHvTNBbRbUCvTNBq-wP1LR26g</recordid><startdate>1997</startdate><enddate>1997</enddate><creator>Leeming, Robert J.</creator><general>Elsevier Science & Technology</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>1997</creationdate><title>Microtiter plate assay for biopterin using cryopreserved Crithidia fasciculata</title><author>Leeming, Robert J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-e262t-42f7233984cdff7f84ecda9aca572ff5513035487cd79298b7a8ef2e67621ef33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Animals</topic><topic>Biological Assay - methods</topic><topic>Biopterins - analogs & derivatives</topic><topic>Biopterins - analysis</topic><topic>Biopterins - biosynthesis</topic><topic>Biopterins - blood</topic><topic>Crithidia fasciculata - growth & development</topic><topic>Cryoprotective Agents</topic><topic>Humans</topic><topic>Molecular Structure</topic><topic>Phenylalanine - blood</topic><topic>Phenylketonurias - metabolism</topic><topic>Reference Values</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Leeming, Robert J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Methods in Enzymology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Leeming, Robert J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Microtiter plate assay for biopterin using cryopreserved Crithidia fasciculata</atitle><jtitle>Methods in Enzymology</jtitle><addtitle>Methods Enzymol</addtitle><date>1997</date><risdate>1997</risdate><volume>281</volume><spage>61</spage><epage>70</epage><pages>61-70</pages><issn>0076-6879</issn><eissn>1557-7988</eissn><isbn>9780121821821</isbn><isbn>012182182X</isbn><abstract>This chapter presents microtiter plate assay for biopterin, using cryopreserved Crithidia fasciculate. The term “total biopterin” includes biopterin, dihydrobiopterin, and tetrahydrobiopterin. Most of the diagnostic criteria and normative data available for the Crithidia assay have been obtained by automated tube assay without chloramphenicol. This requires heating to remove protein, and although heat treatment is not necessary for the microtiter plate assay, the practice has continued for comparative, diagnostic purposes. Plasma samples are diluted 1:30 and whole-blood samples are diluted 1:40 in phosphate buffer. Dried blood spots have an 8-ram circle punched out into 1 ml of phosphate buffer. Samples are then heated in a steamer for 10 min, centrifuged, and passed through 0.2-μm filters to clarify. 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| ispartof | Methods in Enzymology, 1997, Vol.281, p.61-70 |
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| source | ScienceDirect eBooks; MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Biological Assay - methods Biopterins - analogs & derivatives Biopterins - analysis Biopterins - biosynthesis Biopterins - blood Crithidia fasciculata - growth & development Cryoprotective Agents Humans Molecular Structure Phenylalanine - blood Phenylketonurias - metabolism Reference Values Reproducibility of Results Sensitivity and Specificity |
| title | Microtiter plate assay for biopterin using cryopreserved Crithidia fasciculata |
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