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Li T, Guo XH, Wang Y, et al. [Preparation, purification and characterization of the polyclonal antibody against human CMTM4]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008;24(1):41-4
Li, T., Guo, X. H., Wang, Y., Markus, P., Shao, L. N., Song, Q. S., Ma, D. L., & Han, W. L. (2008). [Preparation, purification and characterization of the polyclonal antibody against human CMTM4]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, 24(1), 41-4.
Li, Ting, et al. "[Preparation, purification and characterization of the polyclonal antibody against human CMTM4]." Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology vol. 24,1 (2008): 41-4.
Li T, Guo XH, Wang Y, Markus P, Shao LN, Song QS, Ma DL, Han WL. [Preparation, purification and characterization of the polyclonal antibody against human CMTM4]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jan;24(1):41-4. Chinese. PMID: 18177617.
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Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jan;24(1):41-4.

[Preparation, purification and characterization of the polyclonal antibody against human CMTM4].

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology

[Article in Chinese]
Ting Li, Xiao-huan Guo, Yu Wang, Plate Markus, Lu-ning Shao, Quan-sheng Song, Da-long Ma, Wen-ling Han

Affiliations

  1. Center for Human Disease Genomics, Department of Immunology, School of Basic Medical Sciences, Peking University, Beijing 100083, China. [email protected]

PMID: 18177617

Abstract

AIM: To prepare, purify and characterize the polyclonal antibodies against CMTM4.

METHODS: Four polypeptides named peptide 1, 2, 3, and 4 were synthesized based on the bioinformatics analysis of the three isoforms of CMTM4, CMTM4-v1, -v2, and -v3, and coupled with keyhole limpet hemocyanin (KLH) for immunization. Among them, peptide 1, 2, and 3, common for CMTM4-v1, v2, and v3, were mixed for immunization to prepare the antibody which can recognize all of the three isoforms (Ab1); while peptide 4, which is specific for CMTM4-v1, was injected separately into New Zealand rabbits to prepare the antibody specifically targeting CMTM4-v1 (Ab2). ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, Ab1 and Ab2 were identified by Western blot and immunohistochemistry assays.

RESULTS: The titers of Ab1 and Ab2 were 1:10(5) and 1:10(6), respectively. Western blot confirmed their high specificity. Ab1 could also be used for immunohistochemistry analysis, but Ab2 could not. Immunohistochemistry analysis with Ab1 demonstrated the high expression level of CMTM4 in human testis, which was consistent with the previous result of Northern blot. Moreover, Western blot and immunohistochemistry analysis verified that Ab1 can also recognize mouse Cmtm4.

CONCLUSION: Two specific antibodies against human CMTM4 were obtained, which will be helpful for further study.

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