Preparation of mouse KCTD10 antibody and expression analysis of KCTD10 in neuroepithelium of neural tube and dorsal root ganglion.
- Author:
Ji-Kang SUN
1
;
Bo ZHANG
;
Jian ZHANG
;
Jian-Lin ZHOU
Author Information
1. Key Laboratory of Protein Chemistry and Developmental Biology of Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
genetics;
metabolism;
Antibodies, Monoclonal;
biosynthesis;
genetics;
Carrier Proteins;
immunology;
Embryo, Mammalian;
Epithelium;
metabolism;
Ganglia, Spinal;
metabolism;
Mice;
Neural Tube;
cytology;
metabolism;
Potassium Channels;
biosynthesis;
genetics;
immunology;
Potassium Channels, Voltage-Gated;
biosynthesis;
genetics;
immunology;
Rabbits;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
immunology
- From:
Chinese Journal of Biotechnology
2007;23(6):1011-1016
- CountryChina
- Language:Chinese
-
Abstract:
KCTD10 is a TNF-alpha inducible protein that can interact with the small subunit of DNA polymerase a and PCNA. In order to study the function of KCTD10, we prepared the rabbit anti-mouse KCTD10 polyclonal antibody by using the His-tagged recombinant mouse KCTD10 protein to immune New Zealand white rabbit. Mouse KCTD10 shares significant similarity with PDIP1 (polymerase delta-interacting protein 1) and TNFAIP1 (tumor necrosis factor alpha-induced protein 1) protein,and then KCTD10 polyclonal antiserum possesses cross-reactivity with PDIP1 protein and TNFAIP1 protein. The partially digested fragments of homogeneous proteins PDIP1 and TNFAIP1 were mixed and incubated with anti-KCTD10 antiserum at 4 degrees C for 3 h to deplete unspecific antibodies. Through this method, we removed successfully the cross-reactivity of anti-KCTD10 antibody with PDIP1 and TNFAIP1 and obtained specific anti-KCTD10 antibody. Then, the anti-KCTD10 antibody was used in immunohistochemistry experiments of mouse. The results of immunohistochemistry on whole-mount embryo and paraffin section demonstrated that KCTD10 is highly expressed in neuroepithelium of neural tube and dorsal root ganglion of 12.5 d embryos. These results suggest that KCTD10 may play roles in the development of neuroepithelium of neural tube and dorsal root ganglion.
