[Preparation of mouse KCTD10 antibody and expression analysis of KCTD10 in neuroepithelium of neural tube and dorsal root ganglion].
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.
Sun
Ji-Kang
JK
Key Laboratory of Protein Chemistry and Developmental Biology of Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha.
Zhang
Bo
B
Zhang
Jian
J
Zhou
Jian-Lin
JL
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics; D002352:Carrier Proteins / Q000276:immunology; D004622:Embryo, Mammalian; D004848:Epithelium / Q000378:metabolism; D005727:Ganglia, Spinal / Q000378:metabolism*; D051379:Mice; D054259:Neural Tube / Q000166:cytology / Q000378:metabolism*; D015221:Potassium Channels / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D024642:Potassium Channels, Voltage-Gated / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D043743:Ubiquitin-Protein Ligase Complexes
D000906:Antibodies; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; C543439:KCTD10 protein, mouse; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D011993:Recombinant Fusion Proteins; C499862:PDIP1 protein, mouse; D043743:Ubiquitin-Protein Ligase Complexes