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Generation and characterization of an anti-GP73 monoclonal antibody for immunoblotting and sandwich ELISA

Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellular carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme lin... Full description

Journal Title: Journal of Biomedical Research November 2012, Vol.26(6), pp.467-473
Main Author: Zhang, Aixia
Other Authors: Cao, Brian
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 1674-8301 ; DOI: 10.7555/JBR.26.20120057
Link: https://www.sciencedirect.com/science/article/pii/S1674830112600670
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recordid: elsevier_sdoi_10_7555_JBR_26_20120057
title: Generation and characterization of an anti-GP73 monoclonal antibody for immunoblotting and sandwich ELISA
format: Article
creator:
  • Zhang, Aixia
  • Cao, Brian
subjects:
  • Gp73
  • Monoclonal Antibody
  • Western Blotting
  • Sandwich ELISA
  • Hepatocellular Carcinoma
  • Medicine
ispartof: Journal of Biomedical Research, November 2012, Vol.26(6), pp.467-473
description: Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellular carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent assay (ELISA). Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody (mAb) designated as 6A2 using recombinant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blotting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this...
language: eng
source:
identifier: ISSN: 1674-8301 ; DOI: 10.7555/JBR.26.20120057
fulltext: fulltext
issn:
  • 1674-8301
  • 16748301
url: Link


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titleGeneration and characterization of an anti-GP73 monoclonal antibody for immunoblotting and sandwich ELISA
creatorZhang, Aixia ; Cao, Brian
ispartofJournal of Biomedical Research, November 2012, Vol.26(6), pp.467-473
identifierISSN: 1674-8301 ; DOI: 10.7555/JBR.26.20120057
subjectGp73 ; Monoclonal Antibody ; Western Blotting ; Sandwich ELISA ; Hepatocellular Carcinoma ; Medicine
descriptionRecently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellular carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent assay (ELISA). Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody (mAb) designated as 6A2 using recombinant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blotting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this...
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Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellular carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent assay (ELISA). Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody (mAb) designated as 6A2 using recombinant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blotting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this...

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Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellular carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent assay (ELISA). Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody (mAb) designated as 6A2 using recombinant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blotting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this...

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