Establishment and characterization of calcyclin binding protein (CacyBP) monoclonal antibody.
Anti-calcyclin binding protein (CacyBP) monoclonal antibodies (MAb) were produced using an in vitro immunization method. BALB/c mouse splenocytes were immunized with purified 6 x His-CacyBP fusion protein and fused with myeloma cells using polyethylene glycol (PEG) 4000. By selection using enzyme-linked immunosorbent assay (ELISA), three anti-CacyBP MAbs were obtained. The MAb BD1, whose isotype was IgG1, interacted with the fusion protein. Western blot and immunofluorescence microscopy showed that the MAb BD1 against CacyBP could recognize CacyBP protein derived from human gastric cancer cell lines in both native and denatured forms. This MAb would act as a useful tool for the detection of CacyBP protein in future studies.
Zhai
Huihong
H
State Key Laboratory of Cancer Biology, Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an, PR China.
Shi
Yongquan
Y
Yu
Jinhua
J
Hong
Liu
L
Tang
Hongwei
H
Wang
Jun
J
Hu
Shengjuan
S
Bai
Feihu
F
Fan
Daiming
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013274:Stomach Neoplasms
D000911:Antibodies, Monoclonal; C491778:CACYBP protein, human; D002135:Calcium-Binding Proteins; D011993:Recombinant Fusion Proteins