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Comparison and evaluation of different methods for alpha-MSH labelling.

G Ghanem·F Legros·F Lejeune·J Frühling
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Abstract

We have studied the behaviour of 125I-labelled alpha-MSH under different experimental conditions. Until now, the chloramine T method had been used by most investigators with variable results. We have tested three other labelling techniques based on 125I mild oxidation: (1) an enzymatic method with lactoperoxidase, (2) a sparingly soluble chloramine method (T.D.G.U.) and (3) modified chloramine T procedure, 'the iodine volatilization method'. Labelled hormone obtained after each kind of iodination was assayed for immunoreactivity. In addition, time course degradation was measured by classical RIA incubation procedures. Charcoal-dextran was used to separate bound and free antigen. We have found chloramine T-iodinated alpha-MSH to be significantly more damaged than preparations obtained by other methods and to be less stable when stored at -18 degrees C. No differences were found between the differently labelled 125I-labelled alpha-MSH fresh preparations in binding to surface receptors of human melanoma cell lines in culture.

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AnimalsCattleChloraminesChromatography, GelHumansImmune SeraImmunologic TechniquesIodine RadioisotopesIsotope LabelingLactoperoxidaseMelanocyte-Stimulating HormonesMelanomaRabbitsTosyl CompoundsUreaVolatilizationComparative StudyJournal ArticleResearch Support, Non-U.S. Gov't

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G Ghanem, F Legros, F Lejeune, J Frühling. 1982-10-29. Comparison and evaluation of different methods for alpha-MSH labelling.. https://doi.org/10.1016/0022-1759(82)90063-1

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