Preparation and identification of polyclonal antiserum against heat shock protein 72

OBJECTIVE: To express fusion protein of histamine (His) and human heat shock protein 72 (hHSP72) in Escherichia coli (E. coli), and to prepare hHSP72 antiserum in rabbit

Bibliographische Detailangaben
Veröffentlicht in:Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue. - 1998. - 18(2006), 9 vom: 08. Sept., Seite 531-4
1. Verfasser: Huang, Fan (VerfasserIn)
Weitere Verfasser: Xu, Kang, Wu, Xing-gang, Ma, Zhong-fu, Lin, Zheng
Format: Aufsatz
Sprache:Chinese
Veröffentlicht: 2006
Zugriff auf das übergeordnete Werk:Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue
Schlagworte:Journal Article Research Support, Non-U.S. Gov't HSP72 Heat-Shock Proteins Immune Sera Recombinant Fusion Proteins
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245 1 0 |a Preparation and identification of polyclonal antiserum against heat shock protein 72 
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500 |a Date Completed 14.09.2007 
500 |a Date Revised 09.12.2020 
500 |a published: Print 
500 |a Citation Status MEDLINE 
520 |a OBJECTIVE: To express fusion protein of histamine (His) and human heat shock protein 72 (hHSP72) in Escherichia coli (E. coli), and to prepare hHSP72 antiserum in rabbit 
520 |a METHODS: hHSP72 gene was inserted into pPROEX-1. The recombinant vector was identified by restriction endonuclease digestion analysis and sequence. Fusion protein His-hHSP72 was expressed in E. coli under isopropyl-beta-D-thiogalactoside (IPTG) induction. The rabbit antibody against His-hHSP72 was prepared by using purified His-hHSP72 protein as immunogen, and the specificity and sensitivity of polyclonal antibody were identified by Western blot 
520 |a RESULTS: The restriction endonuclease digestion analysis of recombinant plasmid and sequence demonstrated that the hHSP72 gene had been exactly inserted into pPROEX-1. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the relative molecular mass of the fusion protein was about 73 ku. Western blot result proved that the rabbit polyclonal antiserum could fuse with over 20 ng hHSP72 protein when diluted to 1:100,000 
520 |a CONCLUSION: The polyclonal antibody against hHSP72 can be prepared in E. coli, it is a new reagent with high specificity and sensitivity for the research of hHSP72 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
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700 1 |a Xu, Kang  |e verfasserin  |4 aut 
700 1 |a Wu, Xing-gang  |e verfasserin  |4 aut 
700 1 |a Ma, Zhong-fu  |e verfasserin  |4 aut 
700 1 |a Lin, Zheng  |e verfasserin  |4 aut 
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