A rapid, non-radioactive method for the detection of insulin-like growth factor binding proteins by Western ligand blotting.
Insulin-like growth factor binding proteins (IGFBPs) are produced by many cell types and are present in a wide variety of biological fluids. Detection of IGFBPs in complex fluids is commonly performed by Western ligand blotting using 12SI-IGF-I and/or 125I-IGF-II as the labeling ligand. This method is extremely useful in detecting IGFBPs; however, it requires the use of radioactive materials and usually takes several days to weeks to complete. Herein, we report on a modification of this method using biotinylated IGFs to detect both purified recombinant IGFBPs, as well as IGFBPs in fibroblast conditioned medium. This alternative method has proved sensitive, time- and cost-efficient, and it is environmentally safe.
Fowlkes
J L
JL
Department of Pediatrics, Duke University Medical Center, Durham, North Carolina 27710, USA.
Serra
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D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin; D015153:Blotting, Western / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D006801:Humans; D018969:Insulin-Like Growth Factor Binding Proteins / Q000032:analysis*; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D008024:Ligands; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D013997:Time Factors
D018969:Insulin-Like Growth Factor Binding Proteins; D008024:Ligands; D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D001710:Biotin