Preparation of anti-mouse PGRP-L single-epitope polyclonal antibody / 南方医科大学学报
Journal of Southern Medical University
;
(12): 859-862, 2007.
Artigo
em Chinês
| WPRIM
| ID: wpr-337368
ABSTRACT
<p><b>OBJECTIVE</b>To prepare a single-epitope polyclonal antibody against mouse long peptidoglycan recognition protein (mPGRP-L).</p><p><b>METHODS</b>B cell dominant epitopes of mPGRP-L predicted by bioinformatics were synthesized, and the immunogen was prepared by conjugation of the synthetic peptide and the carrier protein key-hole limpet hemocyanin (KLH) by MBS method. The single-epitope polyclonal antibody was obtained by immunizing rabbits with the KLH-peptide conjugate, purified by SPG affinity columns or antigenic peptide affinity columns, and identified by ELISA and Western blotting.</p><p><b>RESULTS AND CONCLUSION</b>A dominant epitope in N-terminal region of mPGRP-L, with amino acid sequence of NH(2)-(C)DPHSLSPELQALISEVAQHD-COOH, was chosen and synthesized. The titer of anti-serum of the rabbits immunized with the KLH-peptide conjugate was 1256,000. The polyclonal antibody purified with SPG affinity columns and antigenic peptide affinity columns were named as mPGRP-Ln1 and mPGRP-Ln2, respectively. Western blotting demonstrated that the antibody mPGRP-Ln1 could recognize the recombined N-terminal fragment of mPGRP-Ln with a clear band at relative molecular mass of about 29,000, suggesting the successful preparation of the single-epitope polyclonal antibody against the N-terminal region of mPGRP-L.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Ligação Proteica
/
Hemocianinas
/
Ensaio de Imunoadsorção Enzimática
/
Dados de Sequência Molecular
/
Proteínas de Transporte
/
Química
/
Western Blotting
/
Sequência de Aminoácidos
/
Mapeamento de Epitopos
/
Alergia e Imunologia
Limite:
Animais
Idioma:
Chinês
Revista:
Journal of Southern Medical University
Ano de publicação:
2007
Tipo de documento:
Artigo
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