Typing of esterase D by isoelectric focusing

Abstract

staining was negligible when using peroxidase-labelled probes, in contrast to the 1251-labelled antisera where the increased background staining reduced the sensitivity.The use of glutaraldehyde vapor fixation of the immobilized antigens followed by reduction with 0.02 % NaBH4 before performing the immunoassays (71 did not significantly improve the sensitivity, although slightly enhanced staining was observed.The conditions used in these immunoassays may differ from those used by other investigators and may be more or less sensitive compared to others. Therefore, the detection limits do not represent ultimate values, but serve as method to compare the various immunoassays. We conclude that the double sandwich immunoassay is the most attractive approach to detect antibody bound to nitrocellulose immobilized antigen since a) it is most sensitive technique and b), it reduces the number of incubation steps compared to other methods.

Polymorphism of EsD by isoelectric focusing: Description of the new allele EsD*Kofu and phenotyping in bloodstains

“…Our results are superior to the determination limits reported by other workers (Horscroft and Sutton 1983;Budowle 1984;White 1984;Finney et al 1985). With increasing time of storage, the bands of the heterozygous types EsD 2-1, EsD 7-1, and EsD 7-2 became fainter and more indistinct than those of the homozygous types EsD 1 and EsD 2.…”

“…Since isoelectric focusing has recently been introduced for use in the analysis of EsD types, several attempts have been made to demonstrated this enzyme polymorphism from bloodstains (Horscroft and Sutton 1983;Budowle 1984;White 1984;Finney et al 1985;Yuasa et al 1985a). Table 2 summarizes the results for the determination limits of EsD types in bloodstains stored at 37 ° C, room temperature, and 4 ° C obtained by the present isoelectric method.…”

“…Using isoelectric focusing, the EsD*5 allele was later shown to be relatively common in white people, while the EsD*7 allele was reported to occur with appreciable frequency in Japanese by Nishigaki and Itoh (1984). Although these isoelectric focusing methods provided a better separation of the products of EsD*5 or EsD*7 from those of EsD*I and EsD*2, it was rather difficult to distinguish between the products of EsD*I and EsD*2, and several technical improvements have been undertaken to overcome this problem (B~ir et al 1984;Budowle 1984;Divall 1984;Finney et al 1985;Henke et al 1985).…”

Forensic science

“…Isoelectric focusing (IEF) has become a valuable technique in the field of forensic serology, and the use of ultrathin gel is making it even more powerful (335). IEF has been used to type esterase D (ESD) (336)(337)(338), group specific component (Gc) (339-343, 335), erythrocyte acid phosphatase (ACPi) (344), transferrin (Tf) (345), and phosphoglucomutase (PGM) (346)(347)(348)(349)(350)(351)(352)335). The value of ultrathin polyacrylamide gels for the simultaneous separation of PGM and EAP by isoelectric focusing of human red cell lysates and bloodstain extracts has been investigated (353).…”

EsD*5 gene frequency in Tuscany (Italy)

“…EsD*5 is a subdivision of EsD*2. Methods for the simultaneous determination of the three common EsD allozymes (1, 2, and 5) have been described by Kiihnl and Spielmann (1981) and, in the last 2 years, by Budowle (1984), Bfir et al (1984), Divall (1984, and Gradl et al (1985). Better results were obtained by Martin (1981), Olaisen et al (1981), Dykes et al (1982) with IEF: however, their methods could not satisfactorily determine EsD 1, 2-1, and 2 phenotypes, at the same time.…”