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P.8. Isolation and preparation of five human transferrin forms using isoelectric focusing
P.7. Subtyping of haptoglobin followed by i m m u n o - b l o t t i n g
(Hp) by isoelectric focusing technique.
K.Hjalmarsson. State Institute for Blood Group Serology, SRL, Regionsjukhuset, S-581 85 LINKOPING, Sweden In 1985, Teige et al. (1)described a method for subtyping, which is very well adapted for routine cases of disputed paternity. The method will be presented with minor modifications. The technique includes isoelectric focusing of reduced neuraminidase treated serum samples followed by immuno-blotting procedure using antihuman haptoglobin as the first antibody and a peroxidase conjugated second antibody. The Hp band pattern is visualized with hydrogen peroxide and a freshly prepared 4-chloro-l-naphtol solution. 1.Teige B o, Olaisen B. and Pedersen L. Subtyping of haptoglobin Presentation of a new method. Hum. Genet. 70:163-167 (1985).
P.8. Isolation and p r e p a r a t i o n ef five h u m a n forms using isoelectric focusing.
transferrin
S. Petr6n, O. Vesterberg and *H. Jt~rnvall. Chemistry Division, Research Department, National Board of Occupational Safety and Health, S-171 84 SOLNA and *Department of Chemistry I, Karolinska Insfitutet, S-104 01 STOCKHOLM, Sweden. Apart from genetic variation, human transferrin shows another multiplicity and occurs in at least five isoforms with different isoelectric points. With preparative IEF in agarose the five forms were separated and detected as red bands clearly visible in the gel. Up to five bands in gel pieces, taken f r o m different gels but containing the same form, were placed on a new gel and the IEF was repeated. Pure forms were obtained according to analytical IEF and amino acid analysis. Each form was digested with trypsin and analysed by HPLC. The results showed that the differences among the forms were restricted to the glycopeptides, suggesting the presence of variations in carbohydrate parts attached to the transferrin polypeptide.
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(Hp) by isoelectric focusing technique.
K.Hjalmarsson. State Institute for Blood Group Serology, SRL, Regionsjukhuset, S-581 85 LINKOPING, Sweden In 1985, Teige et al. (1)described a method for subtyping, which is very well adapted for routine cases of disputed paternity. The method will be presented with minor modifications. The technique includes isoelectric focusing of reduced neuraminidase treated serum samples followed by immuno-blotting procedure using antihuman haptoglobin as the first antibody and a peroxidase conjugated second antibody. The Hp band pattern is visualized with hydrogen peroxide and a freshly prepared 4-chloro-l-naphtol solution. 1.Teige B o, Olaisen B. and Pedersen L. Subtyping of haptoglobin Presentation of a new method. Hum. Genet. 70:163-167 (1985).
P.8. Isolation and p r e p a r a t i o n ef five h u m a n forms using isoelectric focusing.
transferrin
S. Petr6n, O. Vesterberg and *H. Jt~rnvall. Chemistry Division, Research Department, National Board of Occupational Safety and Health, S-171 84 SOLNA and *Department of Chemistry I, Karolinska Insfitutet, S-104 01 STOCKHOLM, Sweden. Apart from genetic variation, human transferrin shows another multiplicity and occurs in at least five isoforms with different isoelectric points. With preparative IEF in agarose the five forms were separated and detected as red bands clearly visible in the gel. Up to five bands in gel pieces, taken f r o m different gels but containing the same form, were placed on a new gel and the IEF was repeated. Pure forms were obtained according to analytical IEF and amino acid analysis. Each form was digested with trypsin and analysed by HPLC. The results showed that the differences among the forms were restricted to the glycopeptides, suggesting the presence of variations in carbohydrate parts attached to the transferrin polypeptide.
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