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 language | English |
|---|---|
| Pages (from-to) | 176-179 |
| Number of pages | 4 |
| Journal | Journal of Investigative Dermatology |
| Volume | 100 |
| Issue number | 2 |
| Publication status | Published - 1993 Feb |
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ASJC Scopus subject areas
- Dermatology
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Detection of the 170-kDa bullous pemphigoid antigen by immunoprecipitation. / Ebihara, Tamotsu; Hashimoto, Takashi; Kudo, Jun; Gamou, Shinobu; Shimizu, Nobuyoshi; Nishikawa, Takeji.
In: Journal of Investigative Dermatology, Vol. 100, No. 2, 02.1993, p. 176-179.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of the 170-kDa bullous pemphigoid antigen by immunoprecipitation
AU - Ebihara, Tamotsu
AU - Hashimoto, Takashi
AU - Kudo, Jun
AU - Gamou, Shinobu
AU - Shimizu, Nobuyoshi
AU - Nishikawa, Takeji
PY - 1993/2
Y1 - 1993/2
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0027448366&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027448366&partnerID=8YFLogxK
M3 - Article
C2 - 8429239
AN - SCOPUS:0027448366
VL - 100
SP - 176
EP - 179
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
SN - 0022-202X
IS - 2
ER -