Production of anti-peptide antibody of rat brain nitric-oxide synthase

Rong G Zhai, Xing Zu Zhu

Research output: Contribution to journalArticle

Abstract

Aim: To raise antibody of rat brain nitric-oxide synthase (bNOS) through immunizing animal with a peptide of bNOS that can represent the holoprotein. Methods: The amino acid sequence for the bNOS was analyzed by GenePro computer program. According to the hydrophilicity, hydrophobicity, antigenicity, and the potentiality to form protein second structures of α-helix, β-sheet and, β-turn, the structure of bNOS was predicted. The peptide 277-287 was selected that was predicted to be in the antigen epitope of bNOS. The peptide was chemically synthesized, coupled to keyhole limpet hemocyanin carrier protein and injected into rabbits to raise antibody. The specificity of the antibody was tested by enzyme-linked immunosorbent assay, immunohistochemistry, and Western blotting. Results: The antibody bound the protein in rat cerebellum extract. In Western blotting, the antibody bound the protein band of 150 kDa in SDS PAGE, and the binding was inhibited by peptide conjugated with carrier protein. In immunohistochemistry, the stain was collocated with the stain in NADPH dehydrogenase histochemistry. Conclusion: The antibody against the peptide recognized the natural bNOS in rat brain, and the peptide 277-287 was located on the surface of bNOS.

Original languageEnglish
Pages (from-to)204-208
Number of pages5
JournalActa Pharmacologica Sinica
Volume18
Issue number3
StatePublished - May 1 1997
Externally publishedYes

Fingerprint

Anti-Idiotypic Antibodies
Brain
Nitric Oxide Synthase
Peptides
Antibodies
Hydrophobic and Hydrophilic Interactions
Carrier Proteins
Coloring Agents
Western Blotting
Immunohistochemistry
Rats
NADPH Dehydrogenase
Proteins
Antibody Specificity
Immunosorbents
Hydrophilicity
Cerebellum
Hydrophobicity
rat Nos1 protein
Epitopes

Keywords

  • antibodies
  • cerebellum
  • enzyme-linked immunosorbent assay
  • epitopes
  • immunohistochemistry
  • NADPHG dehydrogenase
  • nitric-oxide synthase
  • peptides
  • Western blotting

ASJC Scopus subject areas

  • Chemistry(all)
  • Pharmacology

Cite this

Production of anti-peptide antibody of rat brain nitric-oxide synthase. / Zhai, Rong G; Zhu, Xing Zu.

In: Acta Pharmacologica Sinica, Vol. 18, No. 3, 01.05.1997, p. 204-208.

Research output: Contribution to journalArticle

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