1. A versatile dot-ELISA method with femtomole sensitivity for detecting small peptides.
- Author
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Sithigorngul P, Stretton AO, and Cowden C
- Subjects
- Animals, Antibodies, Monoclonal, Ascaris analysis, Cholecystokinin analysis, Cholecystokinin immunology, Chromatography, High Pressure Liquid, FMRFamide, Invertebrate Hormones analysis, Neuropeptides analysis, Neuropeptides immunology, Rabbits, Sensitivity and Specificity, Enzyme-Linked Immunosorbent Assay methods, Peptides analysis
- Abstract
Several protocols for conjugating peptides in situ to a protein carrier on paper, nitrocellulose, or nylon membranes were explored for their usefulness in dot-ELISA detection of the peptides. The most sensitive method in which peptide diluted in bovine serum albumin is applied to nitrocellulose, then fixed with glutaraldehyde, can detect several peptides, ranging from 4 to 38 amino acids in length, at the level of 2-10 fmol. Both immunohistochemical grade antisera and monoclonal antibodies have been used successfully. The method may be a useful alternative to radioimmunoassay since there is no requirement for radiolabelled peptide, or (for quantitation) for known quantities of unlabelled peptide. The method has been used to monitor, semiquantitatively, the fractionation of FMRFamide-like or CCK-like peptides from the nematode Ascaris, and to detect peptide-like immunoreactivities in tissue extracts.
- Published
- 1991
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