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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
2002-10-31
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pubmed:abstractText |
Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARd and PPARg2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence.Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:issn |
1340-3478
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pubmed:author |
pubmed-author:AoiRieR,
pubmed-author:AuwerxJohanJ,
pubmed-author:DoiTakefumiT,
pubmed-author:HamakuboTakaoT,
pubmed-author:IwanariHirokoH,
pubmed-author:JiangShu-YingSY,
pubmed-author:KodamaTatsuhikoT,
pubmed-author:MangelsdorfDavid JDJ,
pubmed-author:MurakamiTakeshiT,
pubmed-author:NaitoMakotoM,
pubmed-author:ReiterRaphaelR,
pubmed-author:ShulmanAndrew IAI,
pubmed-author:SuzukiAkifumiA,
pubmed-author:TachibanaKeisukeK,
pubmed-author:TakenoTetsuT,
pubmed-author:TanakaToshiyaT,
pubmed-author:UchiyamaYasutoshiY,
pubmed-author:WatanabeYuichiroY,
pubmed-author:YamashitaHisahikoH
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pubmed:issnType |
Print
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pubmed:volume |
9
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
233-42
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pubmed:dateRevised |
2009-11-3
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pubmed:meshHeading |
pubmed-meshheading:12409633-Animals,
pubmed-meshheading:12409633-Antibodies, Monoclonal,
pubmed-meshheading:12409633-Blotting, Western,
pubmed-meshheading:12409633-CHO Cells,
pubmed-meshheading:12409633-Cricetinae,
pubmed-meshheading:12409633-Electrophoretic Mobility Shift Assay,
pubmed-meshheading:12409633-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:12409633-Female,
pubmed-meshheading:12409633-Humans,
pubmed-meshheading:12409633-Immunohistochemistry,
pubmed-meshheading:12409633-Mice,
pubmed-meshheading:12409633-Mice, Inbred BALB C,
pubmed-meshheading:12409633-Nucleopolyhedrovirus,
pubmed-meshheading:12409633-Receptors, Cytoplasmic and Nuclear,
pubmed-meshheading:12409633-Recombinant Fusion Proteins,
pubmed-meshheading:12409633-Spodoptera,
pubmed-meshheading:12409633-Transcription Factors
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pubmed:year |
2002
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pubmed:articleTitle |
The generation of monoclonal antibodies against human peroxisome proliferator-activated receptors (PPARs).
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pubmed:affiliation |
Department of Molecular Biology and Medicine, Research Center for Advanced Science and Technology, The Tokyo University, Tokyo, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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