title: The development of an ELISA for group IVA phospholipase A2 in human red blood cells.
abstract: An immunoassay for IVA phospholipase A2 in human red blood cells is described. The assay is a non-competitive sandwich assay in which increasing amounts of the measured protein produce increased luminescence. The antibodies used in the assay are directed against two unique epitopes of the molecule, which sequentially trap and detect the protein. The standard curve covers the range 0.7ng to 23ng/mL (0.07 to 2.3ng/well). The intra-assay and inter-assay coefficients of variation were 9% and 12%, respectively. Evidence is presented that the assay is specific for the alpha paralog of IV PLA2. The assay allows simple and rapid quantification of IVAPLA2 in red blood cell lysates and other biological fluids.
authors.affiliation: Department of Biochemistry, Victoria Infirmary, Glasgow G42 9TY, UK. Electronic address: donaldjmacdonald760@gmail.com.
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Macdonald
authors.affiliation: Department of Biochemistry, Victoria Infirmary, Glasgow G42 9TY, UK.
authors.forename: Rose M
authors.initials: RM
authors.lastname: Boyle
authors.affiliation: Department of Biochemistry, Victoria Infirmary, Glasgow G42 9TY, UK.
authors.forename: Alastair C A
authors.initials: AC
authors.lastname: Glen
authors.affiliation: Department of Pediatrics, National Jewish Health, Denver, CO 80206, USA.
authors.forename: Christina C
authors.initials: CC
authors.lastname: Leslie
authors.forename: A Iain M
authors.initials: AI
authors.lastname: Glen
authors.forename: David F
authors.initials: DF
authors.lastname: Horrobin
mesh_terms: D000328:Adult; D000818:Animals; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000378:metabolism*; D004912:Erythrocytes / Q000201:enzymology*; D005260:Female; D054513:Group IV Phospholipases A2 / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D061987:Sf9 Cells; D018411:Spodoptera; D055815:Young Adult
chemical_list: D000939:Epitopes; D054513:Group IV Phospholipases A2
keywords: Alpha paralog; Arachidonic acid; Cell signalling; Diagnostic marker; Red blood cell; Sandwich immunoassay