Characterization of Cep85 – a new antagonist of Nek2A that is involved in the regulation of centrosome disjunction | Zendy

Canhe Chen | Zendy; Fang Tian | Zendy; Lin Lü | Zendy; Yun Wang | Zendy; Zhe Xiao | Zendy; Chengtao Yu | Zendy; Xianwen Yu | Zendy

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Characterization of Cep85 – a new antagonist of Nek2A that is involved in the regulation of centrosome disjunction

Author(s) -

Canhe Chen,

Fang Tian,

Lin Lü,

Yun Wang,

Zhe Xiao,

Chengtao Yu,

Xianwen Yu

Publication year - 2015

Publication title -

journal of cell science

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.384

H-Index - 278

eISSN - 1477-9137

pISSN - 0021-9533

DOI - 10.1242/jcs.180463

Subject(s) - biology , transfection , confusion , small interfering rna , sequence (biology) , oligonucleotide , centrosome , microbiology and biotechnology , rna interference , rna , gene , genetics , cell cycle , psychology , psychoanalysis

Fig. S1. Cep85 and rabbit polyclonal antibody anti-Cep85 (A) Schematic diagram of Cep85 protein. coiled-coil (CC) domains, Green: NBD (Nek2A binding domain, identified in this study), aa 257-433; CLD (centrosome localization domain, also identified in this study), aa 434-476. (B) Phylogenetic tree shows that Cep85 is a conserved protein. The percentages represent identities. (C-E) Characterization of Cep85 antibody. (C) Recombinant GST-Cep85 protein with the fragment of aa 544-762 was used as an antigen to immunize rabbits. The indicated Myc-tagged Cep85 proteins were expressed in HEK293T cells and the cell lysates were subjected to Western blotting with antibodies against Myc tag and Cep85. (D) The rabbit anti-Cep85 polyclonal antibody can be used for immunoprecipitation assays. Cell lysate from HEK293T cells overexpressing Myc-Cep85 was subjected to immunoprecipitation (IP) with anti-Cep85 antibody. Immunobotting with anti-Myc antibody recognizes the same spot detected by anti-Cep85 antibody. TCL, total cell lysate.

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