A sandwich enzyme-linked imrmmosorbent assay for Δ3, Δ2-enoyl-CoA isomerase in rat-tissue homogenates

Hideaki Suzuki; Yoshihisa Tomioka; Shunji Ishiwata; Takahumi Yamaguchi; Koichi Miura; Takawori Hishinuma; Michinao Mizugaki

doi:10.1620/tjem.179.35

A sandwich enzyme-linked imrmmosorbent assay for Δ³, Δ²-enoyl-CoA isomerase in rat-tissue homogenates

Hideaki Suzuki, Yoshihisa Tomioka, Shunji Ishiwata, Takahumi Yamaguchi, Koichi Miura, Takawori Hishinuma, Michinao Mizugaki

PHA - Pharmacy

Research output: Contribution to journal › Article › peer-review

Abstract

We established two monoclonal antibodies (MAbs, A6-1-E4, IgM; א and BC-D1, IgG₁, λ), that specifically recognize different epitopes of rat Δ³, Δ²-enoyl-CoA isomerase (ECI). We then developed a specific and reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) using a capture monoclonal antibody and an indicator polyclonal antibody to evaluate ECI. The amounts of ECI were linearly determined in the ranges from 15 to 250 ng/well by this method. This assay can detect the amount of ECI in tissue homogenates without subfractionation. The ECI levels were 0.13 ng/μg homogenate in normal rat liver and 1.50 ng/μg homogenate in clofibrate-treated rat liver.

Original language	English
Pages (from-to)	35-45
Number of pages	11
Journal	Tohoku Journal of Experimental Medicine
Volume	179
Issue number	1
DOIs	https://doi.org/10.1620/tjem.179.35
Publication status	Published - 1996 May

Keywords

Monoclonal antibody
Sandwich ELISA
Δ, Δ-enoyl-CoA-isomerase
β-oxidation of unsaturated fatty acids

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

Access to Document

10.1620/tjem.179.35

Cite this

@article{46659be9d9f440a38f8c45ddfa377c68,

title = "A sandwich enzyme-linked imrmmosorbent assay for Δ3, Δ2-enoyl-CoA isomerase in rat-tissue homogenates",

abstract = "We established two monoclonal antibodies (MAbs, A6-1-E4, IgM; א and BC-D1, IgG1, λ), that specifically recognize different epitopes of rat Δ3, Δ2-enoyl-CoA isomerase (ECI). We then developed a specific and reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) using a capture monoclonal antibody and an indicator polyclonal antibody to evaluate ECI. The amounts of ECI were linearly determined in the ranges from 15 to 250 ng/well by this method. This assay can detect the amount of ECI in tissue homogenates without subfractionation. The ECI levels were 0.13 ng/μg homogenate in normal rat liver and 1.50 ng/μg homogenate in clofibrate-treated rat liver.",

keywords = "Monoclonal antibody, Sandwich ELISA, Δ, Δ-enoyl-CoA-isomerase, β-oxidation of unsaturated fatty acids",

author = "Hideaki Suzuki and Yoshihisa Tomioka and Shunji Ishiwata and Takahumi Yamaguchi and Koichi Miura and Takawori Hishinuma and Michinao Mizugaki",

year = "1996",

month = may,

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T1 - A sandwich enzyme-linked imrmmosorbent assay for Δ3, Δ2-enoyl-CoA isomerase in rat-tissue homogenates

AU - Suzuki, Hideaki

AU - Tomioka, Yoshihisa

AU - Ishiwata, Shunji

AU - Yamaguchi, Takahumi

AU - Miura, Koichi

AU - Hishinuma, Takawori

AU - Mizugaki, Michinao

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N2 - We established two monoclonal antibodies (MAbs, A6-1-E4, IgM; א and BC-D1, IgG1, λ), that specifically recognize different epitopes of rat Δ3, Δ2-enoyl-CoA isomerase (ECI). We then developed a specific and reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) using a capture monoclonal antibody and an indicator polyclonal antibody to evaluate ECI. The amounts of ECI were linearly determined in the ranges from 15 to 250 ng/well by this method. This assay can detect the amount of ECI in tissue homogenates without subfractionation. The ECI levels were 0.13 ng/μg homogenate in normal rat liver and 1.50 ng/μg homogenate in clofibrate-treated rat liver.

AB - We established two monoclonal antibodies (MAbs, A6-1-E4, IgM; א and BC-D1, IgG1, λ), that specifically recognize different epitopes of rat Δ3, Δ2-enoyl-CoA isomerase (ECI). We then developed a specific and reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) using a capture monoclonal antibody and an indicator polyclonal antibody to evaluate ECI. The amounts of ECI were linearly determined in the ranges from 15 to 250 ng/well by this method. This assay can detect the amount of ECI in tissue homogenates without subfractionation. The ECI levels were 0.13 ng/μg homogenate in normal rat liver and 1.50 ng/μg homogenate in clofibrate-treated rat liver.

KW - Monoclonal antibody

KW - Sandwich ELISA

KW - Δ, Δ-enoyl-CoA-isomerase

KW - β-oxidation of unsaturated fatty acids

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