Abstract
The N-terminal fragment of ATP1AL1, the possible catalytic subunit of human ouabain-sensitive H+,K(+)-ATPase, was expressed in Escherichia coli cells as two recombinant proteins: the Ser14-Ile104 fragment or the same fragment containing His6 sequence at its N-end. The second protein was purified by metal-affinity chromatography and used as an antigen to construct two hybridoma lines producing antibodies of the IgM class. These monoclonal antibodies were shown to recognize not only the starting antigen but also the full-size recombinant ATP1AL1 protein and do not react with Na+,K(+)-ATPase.
MeSH terms
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Amino Acid Sequence
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Antibodies, Monoclonal / immunology*
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Cloning, Molecular
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Gene Expression Regulation, Enzymologic / genetics*
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H(+)-K(+)-Exchanging ATPase / biosynthesis
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H(+)-K(+)-Exchanging ATPase / chemistry
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H(+)-K(+)-Exchanging ATPase / genetics*
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H(+)-K(+)-Exchanging ATPase / immunology
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Histidine
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Humans
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Hybridomas / immunology
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Immunoglobulin M / immunology
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Isoleucine
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Molecular Sequence Data
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Recombinant Proteins / genetics
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Serine
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Sodium-Potassium-Exchanging ATPase / chemistry
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Sodium-Potassium-Exchanging ATPase / genetics*
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Sodium-Potassium-Exchanging ATPase / immunology
Substances
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Antibodies, Monoclonal
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Immunoglobulin M
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Recombinant Proteins
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Isoleucine
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Serine
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Histidine
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ATP12A protein, human
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H(+)-K(+)-Exchanging ATPase
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Sodium-Potassium-Exchanging ATPase