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KMID : 1204320090250020145
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Laboratory Animal Research
2009 Volume.25 No. 2 p.145 ~ p.150
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Production and Characterization of Polyclonal Antibody to Transmembrane Inner Ear Protein
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Nam Yoon-Yi
Karuppasamy Sankarapandian
Park Byoung-Kwon
Kwon Hyung-Joo
Suh Jun-Gyo
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Abstract
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Deafness is one of the most common sensory deficits in human. Since the inner ears of mice and humans are anatomically and physiologically similar, mouse mutants allow us to reveal detailed analysis of defects in the cochlea and vestibular apparatus. Loss of function mutations in transmembrane inner ear (Tmie) protein has been shown to cause deafness in mice (circling, spinner) and humans (DFNB6). However, the detailed mechanism of Tmie protein in the cochlea still remains unclear. First step to achieve that purpose, we produced polyclonal antibody that recognize Tmie protein. The specifically selected amino acid sequence localized at the positions 118-133 (IPGEDKKKKKKDSVD) of Tmie protein has been used as immunogen to produce rabbit polyclonal antibody. Two rabbits were immunized with polypeptide and the serum was harvested from the rabbits served as the source of the polyclonal antibody specific to Tmie. Specificity of the antibody was determined by Western blot analysis. After transfection of tmiePC/pcDNA vector in HEK cells, we identified a specific band of tmie protein. Taken together, our anti-Tmie antibody can be useful for studying localization and characterization of Tmie protein.
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KEYWORD
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Tmie, deafness, DFNB6, cochlea, polyclonal antibody
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