PMID: 9166112May 1, 1997Paper

Insulin-like growth factor-I (IGF-I) production by bovine granulosa cells in vitro and peripheral IGF-I measurement in cattle serum: an evaluation of IGF-binding protein extraction protocols

The Journal of Endocrinology
C G GutiérrezR Webb

Abstract

Insulin-like growth factor-binding protein (IGFBP) extraction protocols were tested for their efficacy in removing IGFBPs from bovine plasma and bovine granulosa cell culture medium compared with standard acid exclusion chromatography. Traditional extraction methods, acidification, Sep-Pak, ethanol:acetone:acetic acid (EAA) and EAA-cryoprecipitation (EAA-C), failed to remove all the IGFBPs from both granulosa cell culture medium and plasma. However, EAA and EAA-C treatment of plasma samples did give values similar to those obtained by acid exclusion HPLC, when corrected for extraction efficiency. There was an inverse relationship between insulin-like growth factor-I (IGF-I) concentration in plasma samples, as measured using HPLC chromatography, and IGF-I concentration after EAA extraction. Furthermore, the interference caused by residual IGFBPs differed between samples taken from animals given various treatment that altered peripheral IGF-I concentrations. As for plasma samples, EAA was the most effective extraction method for culture media, but residual IGFBPs caused an overestimation of IGF-I concentrations. In culture media, but not plasma, it was possible to block the interference of IGFBPs in the IGF-I assay, in both extra...Continue Reading

Citations

Jun 24, 2003·Animal Reproduction Science·M Mihm, E C L Bleach
Jul 28, 2009·Reproduction : the Official Journal of the Society for the Study of Fertility·Ali A Fouladi-NashtaRobert Webb
Jul 20, 2007·Reproduction : the Official Journal of the Society for the Study of Fertility·Fiona H ThomasEvelyn E Telfer
Aug 31, 2004·Reproduction : the Official Journal of the Society for the Study of Fertility·E D WatsonD G Armstrong
Jan 9, 2009·Reproduction : the Official Journal of the Society for the Study of Fertility·P C GarnsworthyR Webb
Nov 3, 2007·Reproduction in Domestic Animals = Zuchthygiene·M P GreenM G Hunter
Mar 9, 2007·Biology of Reproduction·Ali A Fouladi-NashtaRobert Webb

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