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Rapid resolution of transferrin C subtypes through isoelectric focusing with 2-mercaptoethanol
Authors:P Auconi  R Donnini  G Bruscalupi  V Pascali
Institution:1. Pediatric Hospital, via Curtatone 54-I52100 Arezzo, Italy;2. Institute of General Physiology, University of Rome Rome, Italy;3. Department of Forensic Medicine, Università Cattolica, Policlinico “Gemelli”, Rome, Italy
Abstract:The technique of choice currently used for the detection of serum transferrin molecular polymorphism is isoelectric focusing on polyacrylamide slab gels. However, this procedure is unsatisfactory for routine purposes, since a long pretreatment of the serum with iron-donor compounds or neuraminidase is necessary in order to obtain a complete resolution of the transferrin molecule. A very fast and highly economical standardized procedure for transferrin typing which enables a fair molecular resolution within only 3 1/2 h is reported. Protracted pretreatment of serum with neuraminidase or with iron-donor compounds can be totally avoided. An ultrathin layer of polyacrylamide gel is employed for the run, using pH ranges of 4-6.5 or 5-7. A short pretreatment of serum with a 13% solution of 2-mercaptoethanol is performed before the samples are placed on the gel. This technique has been used to perform transferrin typing in 396 cord serum samples from newborn infants of Arezzo (Tuscany), without occurrence of artifacts or the appearance of extra bands in transferrin patterns.
Keywords:isoelectric focusing  proteins  To whom correspondence should be addressed: Ospedale Pediatrico  Via Curtatone 54  I52100 Arezzo  Italy  
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