Preparation and characterization of a novel monoclonal antibody specific to human NOB1 protein.
The human NOB1 gene is located at chromosome 16q22.1, encoding a protein consisting of one zinc ribbon domain. Here we aimed to efficiently generate a monoclonal antibody against NOB1 protein. We synthesized the peptide APVEHVVADAGAFLRH based on published NOB1 cDNA sequences. The peptide was chemically linked with the carrier protein keyhole limpet hemocyanin and then injected into Balb/c mice. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using either purified 6xHis-NOB1 fusion protein or the peptide. One MAb named H11 (IgG1), effective in detecting the native NOB1 protein, was characterized by ELISA and Western immunoblotting. By using the MAb, we found NOB1 protein was expressed in human lung, liver, spleen, and kidney tissues. Taken together, the MAb H11 would be helpful for understanding the distribution and functions of NOB1.
Han
Yu
Y
Department of Otolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi Province, China.
Hong
Liu
L
Qiu
Jianhua
J
Qiao
L
L
Zhong
Cuiping
C
Xue
Tao
T
Wang
Menghang
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002874:Chromosome Mapping; D006433:Hemocyanins / Q000737:chemistry; D006825:Hybridomas / Q000378:metabolism; D007158:Immunologic Techniques; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D009687:Nuclear Proteins / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry*; D013154:Spleen / Q000378:metabolism; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; C506664:NOB1 protein, human; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin