Preparation and identification of monoclonal antibody against PNAS-2 protein.
- Author:
Jia LIU
1
;
Fang-Yuan CHEN
;
Hai-Rong WANG
;
Ji-Hua ZHONG
;
Hua ZHONG
;
Jie-Ying HAN
;
Ren-Rong OUYANG
Author Information
1. Department of Hematology, Renji Hospital, Shanghai Jiaotong University Medical College, Shanghai 200001, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
immunology;
isolation & purification;
Antibody Specificity;
immunology;
Apoptosis Regulatory Proteins;
immunology;
Female;
Mice;
Mice, Inbred BALB C
- From:
Journal of Experimental Hematology
2009;17(5):1269-1272
- CountryChina
- Language:Chinese
-
Abstract:
This study was purposed to prepare and primarily identify the specific monoclonal antibodies (McAbs) against the apoptosis related protein PNAS-2 so as to provide the essential tool for study of PNAS-2 function. The McAbs against PNAS-2 were prepared via the immunization of mice, cell fusion and cloning using synthetic peptide of PNAS-2 as immunogen; the specificity, titer and subtype of McAb were detected by Western blot, ELISA and immunofluorescence. The results showed that the stable hybridoma cell line S-31-7 producing McAbs against PNAS-2 protein was successfully obtained. The immunoglobulin of the McAb was identified to be IGg1lambda. The titer of ascetic fluid fled McAb were 1:8,000. A single specific band with 28 kD was shown in Western blot test, and the antigen recognized was present in cell cytoplasm by immunofluorescence. In conclusion, the obtained McAb against PNAS-2 displays strong specificity and high titer, which may be applied to the advanced research on PNAS-2 protein.
