Document Detail


Preparation of polyclonal antibody against human MxA protein and its specificity to diversified myxovirus resistant protein A.
MedLine Citation:
PMID:  20486440     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
OBJECTIVE: To study the human myxovirus resistant protein A (MxA), a specifically induced peptide by interferon I, and to use its level as a diagnostic criterion for viral infections. METHODS: Anti-MxA antisera from immunized mice were prepared with the expressed MxA protein of pET32a-MxA in E. coli BL-21(DE3). To confirm the antiserum activity and specificity, the expression product of BL21, wild type MxA pEGFP-C1-wMxA and site-directed mutant MxA pEGFP-C1-mMxA(N589S) stably transfected 3T3 cells and induced A549 cells were detected by Western blot with the antisera using non-MxA transfected or non-IFN-beta induced cells, intact A549, NIH 3T3 cells transfected with pEGFP-C1 and pET32a (+)-transformed BL-21 as controls. RESULTS: The antisera had specific positive immunoreactivity to the NIH3T3 cells transformed with pEGFP-C1-wMxA and pEGFP-C1-mMxA, INF-beta induced A549 cells and BL21 proteins expressed with pET32a (+)-MxA. The hybridization signals from IFN-beta induced A549 cells depended on the IFN-beta inducing concentrations. Meanwhile, immunohistochemical assay showed that NIH 3T3 cells with pEGFP-C1-wMxA and pEGFP-C1-mMxA had > 98% of positive cells at 1:50 dilution of the serum and A549 cells induced by 20 ng/mL IFN-beta for 48 h showed 95% positive cells. pEGFP-C1-transfected NIH 3T3 cells were all negative. CONCLUSION: Anti-sera are highly specific to diversified MxAs. The antibody is detectable by Western blot, immunocytochemistry and immunofluorescence assay.
Authors:
Kang Wu; Peng Liu; Xiang-Xun Meng; Li Liu; Yan-Hua Li; Zheng-Zhen Ge; Ji-Chen Yang
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biomedical and environmental sciences : BES     Volume:  23     ISSN:  0895-3988     ISO Abbreviation:  Biomed. Environ. Sci.     Publication Date:  2010 Feb 
Date Detail:
Created Date:  2010-05-21     Completed Date:  2010-06-15     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8909524     Medline TA:  Biomed Environ Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  76-82     Citation Subset:  IM    
Affiliation:
School of Basic Medicine and Biology, Suzhou University, Suzhou 215123, Jiangsu, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibody Specificity*
Cell Line, Tumor
GTP-Binding Proteins / genetics,  immunology*,  metabolism
Gene Expression Regulation
Humans
Mice
NIH 3T3 Cells
Species Specificity
Chemical
Reg. No./Substance:
0/myxovirus resistance proteins; EC 3.6.1.-/GTP-Binding Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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