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- W2380340333 abstract "【Objective】 This study was to develop and characterize monoclonal antibodies against Luman-recruiting factor N terminal(LRF-N).【Method】 PCR was used to amplified 194 bp fragment of LRF gene N-terminal and a recombinant plasmid pGEX-LRF-N was constructed.Then,the GST-LRF-N recombinant protein was expressed and purified before being used to immunize the female BALB/c Mus musculus(with age of 6-8 weeks) once every 10 days for a total of 4 times.The spleens were extracted 3 days after the fourth immunization,and splenocytes were fused with SP2/0 cells.The selection and identification of positive samples and the identification of karyotypes,secretion stabilities and the obtained 3 monoclonal antibodies against GST-LRF-N(MAb,named as 2AC9,2AG10,and 3G7,respectively) were finally conducted by indirect ELISA and Western blot assays.【Result】 Recombinant plasmid pGEX-LRF-N was expressed efficiently in the Escherichia coli BL21(DE3),and the molecular weight of its expressive GST-LRF-N protein was 33 ku,Mouse positive antiserum could recognize GST-LRF-N protein.The MAbs 2AC9,2AG10,and 3G7 could specifically recognize pGEX-LRF-N,from pGEX-Luman protein,pGEX-Atac2 protein and pGEX-4T-1 protein.The results of isotype analysis showed that MAbs 2AC9,2AG10,and 3G7 could be categorized to IgG2a,IgM,and IgG1 subclasses,respectively.【Conclusion】 The obtained MAbs from 2AC9,2AG10,and 3G7 hybridoma cells have good monospecificities and can stably secrete antibodies,thus can be used for further research of LRF protein in mouse tissues." @default.
- W2380340333 created "2016-06-24" @default.
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- W2380340333 date "2012-01-01" @default.
- W2380340333 modified "2023-09-24" @default.
- W2380340333 title "Preparation and identification of monoclonal antibodies against Luman-recruiting factor N-terminal" @default.
- W2380340333 hasPublicationYear "2012" @default.
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