Summary
A simple isoelectric focusing method for haptoglobin (HP) typing is described. Serum was pretreated first with C. perfringens neuraminidase (CPN) and then with dithiothreitol (DTT). The treated serum was subjected to polyacrylamide gel isoelectric focusing (PAGIF), and the band patterns were detected by immunoblotting. The method could be successfully applied to HP typing of bloodstains as old as 2 months. A slight modification of it enabled HP, complement component C81, and factor I (IF) to be typed simultaneously. The immunoblotting facilitated preservation of HP patterns. Thus, the PAGIF method for HP typing is suitable for routine use in the forensic laboratory.
Zusammenfassung
Es wird eine einfache Methode zur Typisierung von Haptoglobin (HP) mittels Isoelektrofokussierung beschrieben. Das Serum wird mit Clostridium perfringens Neuraminidase (CPN), sodann mit Dithiothreitol (DTT) vorbehandelt und anschließend der Polyacrylamidgel-Isoelektrofokussierung (PAGIF) unterworfen. Die Banden wurden durch Immunoblotting sichtbar gemacht. Die Methode erwies sich als erfolgreich bei der HP-Darstellung von selbst 2 Monate alten Blutspuren. Durch eine leichte Modifizierung ist es möglich, neben HP auch die Komplement-Komponente C81 und Faktor I (IF) darzustellen. Immunoblotting ergab außerdem eine dauerhafte Darstellung der HP-Banden. PAGIF ist daher als forensische Routinemethode zur HP-Typisierung geeignet.
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Supported in part by Grant-in-Aid for Encouragement of Young Scientest No. 62770408 from The Ministry of Education, Science and Culture
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Fukuda, M., Tamaki, Y. & Kishida, T. Haptoglobin phenotyping by polyacrylamide gel isoelectric focusing and its application to simultaneous typing of serum proteins. Z Rechtsmed 101, 37–40 (1988). https://doi.org/10.1007/BF00205322
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DOI: https://doi.org/10.1007/BF00205322