Detection of the 170-kDa bullous pemphigoid antigen by immunoprecipitation

Tamotsu Ebihara, Takashi Hashimoto, Jun Kudoh, Shinobu Gamou, Nobuyoshi Shimizu, Takeji Nishikawa

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12 Citations (Scopus)

Abstract

There has been controversy concerning the nature of the bullous pemphigoid (BP) antigen: immunoprecipitation identified BP antigen as a single, unique 230-kDa protein, whereas immunoblot analysis showed multiple antigen molecules, mainly 230- and 170-kDa proteins. In this study, to further characterize the 170-kDa protein, we have examined whether the 170-kDa protein is detected by immunoprecipitation. Extracts of human squamous cell carcinoma cells revealed the 170-kDa protein with immunoblot analysis. Although the conventional immunoprecipitation detected only the 230-kDa protein, some BP sera that detected the 170-kDa protein with immunoblotting also precipitated the 170-kDa protein with our modified immunoprecipitation, in which the cells were extracted with 1% sodium dodecylsulfate (SDS) buffer and reacted with the sera under reduced SDS concentration. The 170-kDa protein-specific BP sera clearly showed hemidesmosomal plaque staining with immunofluorescence of cultured cells. These results indicate that the 170-kDa protein is indeed one of the BP antigens and that the 230- and 170-kDa BP antigens are integrated in different ways in hemidesmosomes.

Original languageEnglish
Pages (from-to)176-179
Number of pages4
JournalJournal of Investigative Dermatology
Volume100
Issue number2
DOIs
Publication statusPublished - 1993 Feb

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology
  • Cell Biology

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    Ebihara, T., Hashimoto, T., Kudoh, J., Gamou, S., Shimizu, N., & Nishikawa, T. (1993). Detection of the 170-kDa bullous pemphigoid antigen by immunoprecipitation. Journal of Investigative Dermatology, 100(2), 176-179. https://doi.org/10.1111/1523-1747.ep12462799