[Preparation and identification of monoclonal antibody against PNAS-2 protein].
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.
Liu
Jia
J
Department of Hematology, Renji Hospital, Shanghai Jiaotong University Medical College, Shanghai 200001, China.
Chen
Fang-Yuan
FY
Wang
Hai-Rong
HR
Zhong
Ji-Hua
JH
Zhong
Hua
H
Han
Jie-Ying
JY
Ouyang
Ren-Rong
RR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity / Q000276:immunology; D051017:Apoptosis Regulatory Proteins / Q000276:immunology*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D051017:Apoptosis Regulatory Proteins