Enzyme-Linked Immunoassay for Midkine, and Its Application to Evaluation of Midkine Levels in Developing Mouse Brain and Sera from Patients with Hepatocellular Carcinomas.
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概要
- 論文の詳細を見る
Midkine (MK) is a growth factor that promotes neurite outgrowth and survival of neurons, and enhances the plasminogen activator in endothelial cells. A highly sensitive enzymelinked immunoassay for MK was developed, involving affinity-purified anti-MK antibodies, their biotinylated form, and avidin-β-galactosidase. The amount of bound avidin-β-galactosidase was determined using a fluorogenic substrate, 4-methylumbelliferyl-β-D-galactoside. This method allowed the detection of human and mouse MK in the range of 50pg-10ng. Pleiotrophin, which is related to MK in its amino acid sequence, did not show any cross reactivity. Employing this method, the MK levels in the developing mouse brain were determined. The MK level was 2μg/g of wet tissue on the 12th day of gestation, and then steadily decreased during embryogenesis and postnatal development to 30ng/g two months after birth. The assay method can also be applied to serum samples. Although the MK levels in the sera of normal human subjects were low or undetectable, 0.6-8ng/ml of MK was detected in samples in the majority of cases of hepatocellular carcinomas.
- 社団法人 日本生化学会の論文
著者
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Inui Tatsuya
Peptide Institute Inc. Protein Research Foundation
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Song Xiao-jun
Department Of Biochemistry Nagoya University School Of Medicine
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Kadomatsu Kenji
Department Of Biochemistry Nagoya University Graduate School Of Medicine
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Hada Hajime
Department Of Internal Medicine Okayama University School Of Medicine
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Muramatsu Hisako
Department Od Biochemistry Nagoya University School Of Medicine
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Tsuji Takao
Department Of Cardiovascular Medicine Okayama University Graduate School Of Medicine And Dentistry
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Koide Norio
Department Of Internal Medicine Okayama University School Of Medicine
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MURAMATSU Takashi
Department of Animal Breeding and Reproduction, Faculty of Agriculture Utsunomiya University
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Sakakibara Shumpei
Peptide Center, Institute for Protein Research, Osaka University
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Kimura Terutoshi
Peptide Center, Institute for Protein Research, Osaka University
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