Antipeptide antibodies against the 5-HT1A receptor

Efrain C. Azmitia; Ilje Yu; Homayoon M. Akbari; Nancy Kheck; Patricia M. Whitaker-Azmitia; Daniel R. Marshak

doi:10.1016/0891-0618(92)90016-J

Antipeptide antibodies against the 5-HT_1A receptor

Efrain C. Azmitia, Ilje Yu, Homayoon M. Akbari, Nancy Kheck, Patricia M. Whitaker-Azmitia, Daniel R. Marshak

Biology and Genomics

Research output: Contribution to journal › Article › peer-review

Abstract

The availability of the primary amino acid sequence for a large numbers of molecules provides a fruitful opportunity for their cellular localization by utilizing the procedure of antipeptide antibody formation. This procedure permits a synthetic peptide sequence to be attached to a carrier molecule for the purpose of inoculating an animal to raise specific antibodies against the selected protein sequence. In this report we describe a number of steps that can be taken to increase the likelihood that the selected peptide sequence will be specific and antigenic. In addition, we describe how the peptides are synthesized, purified and coupled to keyhold limpet hemocyanin. The preparation of the antibody and its characterization are also presented in this method report. The immunocytochemical staining at both the light and ultrastructural level with serotonin (5-HT_1A) receptor antipeptide antibodies is discussed. The advantages and disadvantages of this procedure are summarized.

Original language	English (US)
Pages (from-to)	289-298
Number of pages	10
Journal	Journal of chemical neuroanatomy
Volume	5
Issue number	4
DOIs	https://doi.org/10.1016/0891-0618(92)90016-J
State	Published - 1992

Keywords

Immunocytochemistry
Serotonin
Ultrastructure

ASJC Scopus subject areas

Cellular and Molecular Neuroscience

Access to Document

10.1016/0891-0618(92)90016-J

Cite this

@article{7907e8a70e394931b4d45f63254a3dac,

title = "Antipeptide antibodies against the 5-HT1A receptor",

abstract = "The availability of the primary amino acid sequence for a large numbers of molecules provides a fruitful opportunity for their cellular localization by utilizing the procedure of antipeptide antibody formation. This procedure permits a synthetic peptide sequence to be attached to a carrier molecule for the purpose of inoculating an animal to raise specific antibodies against the selected protein sequence. In this report we describe a number of steps that can be taken to increase the likelihood that the selected peptide sequence will be specific and antigenic. In addition, we describe how the peptides are synthesized, purified and coupled to keyhold limpet hemocyanin. The preparation of the antibody and its characterization are also presented in this method report. The immunocytochemical staining at both the light and ultrastructural level with serotonin (5-HT1A) receptor antipeptide antibodies is discussed. The advantages and disadvantages of this procedure are summarized.",

keywords = "Immunocytochemistry, Serotonin, Ultrastructure",

author = "Azmitia, {Efrain C.} and Ilje Yu and Akbari, {Homayoon M.} and Nancy Kheck and Whitaker-Azmitia, {Patricia M.} and Marshak, {Daniel R.}",

note = "Funding Information: We would like to thank G. Binns and M. Meneilly for the preparation of the peptides at Cold Spring Harbor Laboratory Protein Chemistry Core Facility, and Lisa Bianco for preparing the antibody at Cold Spring Harbor Laboratory Animal Facility. Help was provided by Y. Chert in light microscopy and by P. Gannon in ultrastructural immunocytochemical analysis of the antibody. We thank Dr R. Kobayashi for his biochemical expertise and Dr J. Traber for his sponsorship. The work was supported by the NSF DevelopmentalN euroscienceS ectiona nd a generous gift from Miles Inc.",

year = "1992",

doi = "10.1016/0891-0618(92)90016-J",

language = "English (US)",

volume = "5",

pages = "289--298",

journal = "Journal of Chemical Neuroanatomy",

issn = "0891-0618",

publisher = "Elsevier",

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T1 - Antipeptide antibodies against the 5-HT1A receptor

AU - Azmitia, Efrain C.

AU - Yu, Ilje

AU - Akbari, Homayoon M.

AU - Kheck, Nancy

AU - Whitaker-Azmitia, Patricia M.

AU - Marshak, Daniel R.

N1 - Funding Information: We would like to thank G. Binns and M. Meneilly for the preparation of the peptides at Cold Spring Harbor Laboratory Protein Chemistry Core Facility, and Lisa Bianco for preparing the antibody at Cold Spring Harbor Laboratory Animal Facility. Help was provided by Y. Chert in light microscopy and by P. Gannon in ultrastructural immunocytochemical analysis of the antibody. We thank Dr R. Kobayashi for his biochemical expertise and Dr J. Traber for his sponsorship. The work was supported by the NSF DevelopmentalN euroscienceS ectiona nd a generous gift from Miles Inc.

PY - 1992

Y1 - 1992

N2 - The availability of the primary amino acid sequence for a large numbers of molecules provides a fruitful opportunity for their cellular localization by utilizing the procedure of antipeptide antibody formation. This procedure permits a synthetic peptide sequence to be attached to a carrier molecule for the purpose of inoculating an animal to raise specific antibodies against the selected protein sequence. In this report we describe a number of steps that can be taken to increase the likelihood that the selected peptide sequence will be specific and antigenic. In addition, we describe how the peptides are synthesized, purified and coupled to keyhold limpet hemocyanin. The preparation of the antibody and its characterization are also presented in this method report. The immunocytochemical staining at both the light and ultrastructural level with serotonin (5-HT1A) receptor antipeptide antibodies is discussed. The advantages and disadvantages of this procedure are summarized.

AB - The availability of the primary amino acid sequence for a large numbers of molecules provides a fruitful opportunity for their cellular localization by utilizing the procedure of antipeptide antibody formation. This procedure permits a synthetic peptide sequence to be attached to a carrier molecule for the purpose of inoculating an animal to raise specific antibodies against the selected protein sequence. In this report we describe a number of steps that can be taken to increase the likelihood that the selected peptide sequence will be specific and antigenic. In addition, we describe how the peptides are synthesized, purified and coupled to keyhold limpet hemocyanin. The preparation of the antibody and its characterization are also presented in this method report. The immunocytochemical staining at both the light and ultrastructural level with serotonin (5-HT1A) receptor antipeptide antibodies is discussed. The advantages and disadvantages of this procedure are summarized.

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