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Immunoblot observation of antigenic protein fractions in Paragonimus westermani reacting with human patients sera
Sung Hwan Kim,Yoon Kong,Suk Il Kim,Shin Yong Kang and Seung-Yull Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract
In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis (SDS-PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblotted. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable.
Figures
Fig. 1 Finding of SDS-PAGE of saline extract of adult P. westermani. SDS-PAGE was done in 10~15% linear gradient gel. Antigen was heat-treated with sample buffer containing 10% 2-mercaptoethanol. Coomassie brilliant blue R-250 stained.
Mr: Molecular weight of marker proteins in kDa.
BPB: Dye front of bromphenol blue.
Fig. 2 SDS-PAGE/ immunoblot finding in 20 active human paragonimiasis patients.
Mr: Molecular weight in kDa.
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