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タイトル: A time- and cost-saving method of producing rat polyclonal antibodies
著者: Wakayama, Tomohiko
Kato, Yukio link image link image
Utsumi, Rie
Tsuji, Akira link image link image
Iseki, Shoichi link image
若山, 友彦
加藤, 将夫
辻, 彰
井関, 尚一
ワカヤマ, トモヒコ
発行日: 2006年 7月
出版社(者): Japan Society of Histochemistry and Cytochemistry = 日本組織細胞化学会
雑誌名: Acta Histochemica et Cytochemica
ISSN: 0044-5991
巻: 39
号: 3
開始ページ: 79
終了ページ: 87
キーワード: Immunohistochemistry
Rat polyclonal antibodies
Recombinant oligopeptides
Wistar rat
抄録: Producing antibodies usually takes more than three months. In the present study, we introduce a faster way of producing polyclonal antibodies based on preparation of the recombinant oligopeptide as antigen followed by immunization of rats. Using this method, we produced antisera against two mouse proteins: ERGIC-53 and c-Kit. An expression vector ligated with a pair of complementary synthetic oligodeoxyribonucleotides encoding the protein was introduced into bacteria, and the recombinant oligopeptide fused with the carrier protein glutathione-S-transferase was purified. Wistar rats were immunized by injecting the emulsified antigen subcutaneously into the hind footpads, followed by a booster injection after 2 weeks. One week after the booster, the sera were collected and examined for the antibody titer by immunohistochemistry. Antisera with 1600-fold titer at the maximum were obtained for both antigens and confirmed for their specificity by Western blotting. Anti-ERGIC-53 antisera recognized acinar cells in the sublingual gland, and anti-c-Kit antisera recognized spermatogenic and Leydig cells in the testis. These antisera were applicable to fluorescent double immunostaining with mouse monoclonal or rabbit polyclonal antibodies. Consequently, this method enabled us to produce specific rat polyclonal antisera available for immunohistochemistry in less than one month at a relatively low cost. © 2006 The Japan Society of Histochemistry and Cytochemistry.
DOI: 10.1267/ahc.06003
URI: http://hdl.handle.net/2297/19338
関連URI: http://www.jstage.jst.go.jp/article/ahc/39/3/39_79/_article
資料種別: Journal Article
権利関係: Copyright (c) 2006 By the Japan Society of Histochemistry and Cytochemistry / 日本組織細胞化学会の許諾を得て登録
版表示: publisher
出現コレクション:1. 査読済論文(医学・保健)

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