Preparation and characterization of a specific monoclonal antibody against CIAPIN1.
Cytokine-induced antiapoptosis inhibitor 1 (CIAPIN1) is a newly identified antiapoptosis molecule and a mediator of gastric MDR. We cloned cDNA of human CIAPIN1 by RT-PCR and constructed prokaryotic expression vectors of human CIAPIN1 by inserting human CIAPIN1 coding region into pET28-a(+) and pGEX- 4T-1, respectively. The fusion proteins were expressed in Escherichia coli and purified by affinity chromotography. Monoclonal antibody (MAb) against CIAPIN1 was obtained with standard cell fusion technique and ELISA screening. Immunohistochemistry and Western blot showed that the anti-CIAPIN1 MAb recognizes human and mouse CIAPIN1 protein in both native and denatured form. Western blotting confirmed that the expression of CIAPIN1 was upregulated in MDR gastric cancer cell lines. This MAb will be a useful tool for the detection of CIAPIN1 protein in future studies.
Hao
Zhiming
Z
State Key Laboratory of Cancer Biology, Department of Gastroenterology, Xijing Hospital, Fourth Military Medical University, Xi'an, People's Republic of China.
Qiao
Taidong
T
Jin
Xiaohang
X
Li
Xiaohua
X
Gao
Juan
J
Fan
Daiming
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D017209:Apoptosis; D015153:Blotting, Western; D002459:Cell Fusion; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D016207:Cytokines; D018076:DNA, Complementary; D018432:Drug Resistance, Multiple / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D006825:Hybridomas; D007114:Immunization; D007150:Immunohistochemistry; D007377:Interleukin-3 / Q000276:immunology*; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; C489114:CIAPIN1 protein, human; D016207:Cytokines; D018076:DNA, Complementary; D007377:Interleukin-3; D047908:Intracellular Signaling Peptides and Proteins; D011993:Recombinant Fusion Proteins