A time- and cost-saving method of producing rat polyclonal antibodies

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概要

• 論文の詳細を見る

• 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.

• 日本組織細胞化学会の論文

著者

• Tsuji Akira

  金沢大学 自然科学研究科創剤科学

• Iseki Shoichi

  Kanazawa Univ. Kanazawa Jpn

• Iseki Shoichi

  Department Of Histology And Embryology Graduate School Of Medical Science Kanazawa University

• Iseki Shoichi

  Department Of Anatomy Kanazawa University School Of Medicine

• Wakayama Tomohiko

  Kanazawa Univ. Kanazawa Jpn

• Wakayama Tomohiko

  Department Of Histology And Embryology Graduate School Of Medical Science Kanazawa University

• Wakayama T

  Department Of Anatomy And Reproductive Cell Biotogy Miyazaki Medicat Couege

• Wakayama Tomohiko

  宮崎医科大学 第1解剖

• Wakayama Tomohiko

  金沢大学 自然科学研究科薬物動態学

• KATO Yukio

  Division of Pharmaceutical Sciences, Graduate School of Natural Science & Technology

• TSUJI Akira

  Division of Pharmaceutical Sciences, Graduate School of Natural Science & Technology

• Utsumi Rie

  Division of Pharmaceutical Sciences, Graduate School of Natural Science and Technology, Kanazawa Uni

• Utsumi Rie

  Division Of Pharmaceutical Sciences Graduate School Of Natural Science And Technology Kanazawa Unive

• Wakayama Tomohiko

  Department Of Anatomy And Reproductive Cell Biotogy Miyazaki Medicat Couege

• Wakayama Tomohiko

  Department Of Anatomy School Of Medicine Kanazawa Univeristy

• Tsuji A

  金沢大学 自然科学研究科創剤科学

• Kubo Yoshiyuki

  金沢大学 自然科学研究科創剤科学

• Kato Yukio

  Division Of Life Science Graduate School Of Agriculture Meiji University

• Tsuji Akira

  Division Of Pharmaceutical Sciences Graduate School Of Natural Science And Technology Kanazawa Unive

• Kato Yukio

  Department Of Dental And Medical Biochemistry Hiroshima University Graduate School Of Biomedical Sci

• Kato Yukio

  金沢大学 自然科学研究科創剤科学

• Iseki Shoich

  Department of Histology and Embryology, Graduate School of Medical Science, Kanazawa University

• Kubo Yoshiyuki

  Faculty Of Pharmacy Institute Of Medical Pharmaceutical And Health Sciences Graduate School Of Medic

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