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用于初步鉴定GPATCH1(一种与人类骨质疏松症相关的新型剪接因子)的多克隆抗体的研制

Development of Polyclonal Antibodies for the Preliminary Characterization of GPATCH1, a Novel Splicing Factor Associated with Human Osteoporosis.

作者信息

Abudukeremu Aikedaimu, Azatibieke Guliqiati, Yimiti Gulisitan, Guan Yaqun, Chen Zhe

机构信息

State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Xinjiang Key Laboratory of Molecular Biology for Endemic Diseases, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xinjiang Medical University, Urumqi, 830017, Xinjiang, China.

出版信息

Appl Biochem Biotechnol. 2025 Mar;197(3):1790-1804. doi: 10.1007/s12010-024-05132-w. Epub 2024 Nov 28.

Abstract

Specific antibodies, which can be used in various experiments, are critical tools for unraveling genes' function, but many commercial antibodies are not tested for these properties. GPATCH1 is a novel G-patch family protein. Genome-wide association studies (GWAS) revealed it as a gene associated with human osteoporosis, and yeast-based research suggested it may be a splicing factor; however, its molecular mechanism remains a mystery. We report here that currently available commercial GPATCH1 antibodies have poor specificity and are not recommended for immunoprecipitation. We elucidated the apparent molecular weight of GPATCH1 to evaluate the antibodies' specificity. Based on this, a specific polyclonal antibody against GPATCH1 that can be used for Western blotting, immunoprecipitation and immunofluorescence was prepared. With the antibodies, we found that GPATCH1 may be a tissue-specific splicing factor. Our study lays the groundwork for further investigations into the molecular mechanisms by which GPATCH1 affects bone metabolism in the future.

摘要

特异性抗体可用于各种实验,是揭示基因功能的关键工具,但许多商业抗体并未针对这些特性进行测试。GPATCH1是一种新型的G-补丁家族蛋白。全基因组关联研究(GWAS)表明它是一种与人类骨质疏松症相关的基因,基于酵母的研究表明它可能是一种剪接因子;然而,其分子机制仍然是个谜。我们在此报告,目前可用的商业GPATCH1抗体特异性较差,不建议用于免疫沉淀。我们阐明了GPATCH1的表观分子量以评估抗体的特异性。在此基础上,制备了一种可用于蛋白质免疫印迹、免疫沉淀和免疫荧光的针对GPATCH1的特异性多克隆抗体。利用这些抗体,我们发现GPATCH1可能是一种组织特异性剪接因子。我们的研究为未来进一步研究GPATCH1影响骨代谢的分子机制奠定了基础。

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