title: [Preparation and application of mouse anti-AEG-1 monoclonal antibody].
abstract: Objective To prepare the fluorescein isothiocyanate (FITC)-labeled mouse monoclonal antibody (mAb) against astrocyte elevated gene-1 (AEG-1) and evaluate its binding ability to tumor cells. Methods The mouse anti-AEG-1 mAb was purified from mouse ascites. The purity and activity of the mAb were identified by SDS-PAGE, Western blotting and immunohistochemistry (IHC). The FITC labeling efficiency was examined by flow cytometry (FCM) and the binding ability of FITC-labeled mAb to tumor cells was detected by FCM and IHC. Results The AEG-1 mAb was successfully purified form mouse ascites, and successfully labeled by FITC. FCM showed that the fluorescence intensity of tumor cells bound with FITC- mAb was significantly strengthened, and the binding rate reached more than 90%. Moreover, the FITC-labeled anti-AEG-1 mAb could specifically recognize and bind to AEG-1 positive tumor cells. Conclusion The mouse anti-AEG-1 mAb has been successfully purified and labeled by FITC, which can specifically bind to tumor cells in vivo and in vitro.
authors.affiliation: Department of Clinical Laboratory, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors.forename: Man
authors.initials: M
authors.lastname: Lu
authors.affiliation: Department of Clinical Laboratory, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors.forename: Min
authors.initials: M
authors.lastname: Long
authors.affiliation: Department of Clinical Laboratory, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Department of Clinical Laboratory, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Clinical Laboratory, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. *Corresponding author, E-mail: huizz328@163.com.
authors.forename: Huizhong
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism*; D001201:Ascites / Q000378:metabolism; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism; D051379:Mice; D016601:RNA-Binding Proteins
chemical_list: D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; C483038:Mtdh protein, mouse; D016601:RNA-Binding Proteins; D016650:Fluorescein-5-isothiocyanate