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 language | English |
|---|---|
| Pages (from-to) | 204-208 |
| Number of pages | 5 |
| Journal | Acta Pharmacologica Sinica |
| Volume | 18 |
| Issue number | 3 |
| State | Published - May 1 1997 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Production of anti-peptide antibody of rat brain nitric-oxide synthase
AU - Zhai, Rong G
AU - Zhu, Xing Zu
PY - 1997/5/1
Y1 - 1997/5/1
N2 - 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.
AB - 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.
KW - antibodies
KW - cerebellum
KW - enzyme-linked immunosorbent assay
KW - epitopes
KW - immunohistochemistry
KW - NADPHG dehydrogenase
KW - nitric-oxide synthase
KW - peptides
KW - Western blotting
UR - http://www.scopus.com/inward/record.url?scp=0030950315&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030950315&partnerID=8YFLogxK
M3 - Article
C2 - 10072933
AN - SCOPUS:0030950315
VL - 18
SP - 204
EP - 208
JO - Zhongguo yao li xue bao = Acta pharmacologica Sinica
JF - Zhongguo yao li xue bao = Acta pharmacologica Sinica
SN - 0253-9756
IS - 3
ER -