人细胞系和原代组织中己糖激酶3蛋白的可靠检测。

IF 2.1 4区 生物学 Q4 CELL BIOLOGY
European Journal of Histochemistry Pub Date : 2025-01-21 Epub Date: 2025-03-12 DOI:10.4081/ejh.2025.4175
Yasmeen H Mady, Carmen G Kalbermatter, Maarij Khan, Anna M Schläfli, Rina Mehmeti, Inti Zlobec, Lucine Christe, Mario P Tschan
{"title":"人细胞系和原代组织中己糖激酶3蛋白的可靠检测。","authors":"Yasmeen H Mady, Carmen G Kalbermatter, Maarij Khan, Anna M Schläfli, Rina Mehmeti, Inti Zlobec, Lucine Christe, Mario P Tschan","doi":"10.4081/ejh.2025.4175","DOIUrl":null,"url":null,"abstract":"<p><p>Accurate differentiation of homologous proteins that share high sequence identity remains a significant challenge in biomedical research, as conventional antibodies often lack sufficient specificity, leading to potential misinterpretations. This issue is particularly evident in the study of hexokinases, a family of isoenzymes that catalyze the first step of glycolysis by phosphorylating glucose. Beyond their canonical metabolic roles, hexokinases play critical non-glycolytic functions, especially in cancer biology. However, their unique tissue distributions and context-dependent roles are often obscured by the overlapping specificities of commercially available antibodies, which can produce misleading results. In this study, we rigorously evaluated a panel of antibodies targeting hexokinase isoenzyme 3 (HK3), highlighting the widespread issue of cross-reactivity and insufficient validation. Through this process, we identified and validated a highly specific antibody for HK3, demonstrating its reliability in western blot and immunohistochemistry applications. Using this validated tool, we reveal the distinct localization of HK3 in myeloid cell populations, providing new insights into its potential functional roles in these cells. This work addresses a critical gap in antibody specificity and establishes HK3 as a uniquely expressed gene in myeloid and immune cells and is absent in other cell types under basal conditions. Providing a foundation for future investigations into its context-dependent functions.</p>","PeriodicalId":50487,"journal":{"name":"European Journal of Histochemistry","volume":"69 1","pages":""},"PeriodicalIF":2.1000,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11956552/pdf/","citationCount":"0","resultStr":"{\"title\":\"Reliable hexokinase 3 protein detection in human cell lines and primary tissue.\",\"authors\":\"Yasmeen H Mady, Carmen G Kalbermatter, Maarij Khan, Anna M Schläfli, Rina Mehmeti, Inti Zlobec, Lucine Christe, Mario P Tschan\",\"doi\":\"10.4081/ejh.2025.4175\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Accurate differentiation of homologous proteins that share high sequence identity remains a significant challenge in biomedical research, as conventional antibodies often lack sufficient specificity, leading to potential misinterpretations. This issue is particularly evident in the study of hexokinases, a family of isoenzymes that catalyze the first step of glycolysis by phosphorylating glucose. Beyond their canonical metabolic roles, hexokinases play critical non-glycolytic functions, especially in cancer biology. However, their unique tissue distributions and context-dependent roles are often obscured by the overlapping specificities of commercially available antibodies, which can produce misleading results. In this study, we rigorously evaluated a panel of antibodies targeting hexokinase isoenzyme 3 (HK3), highlighting the widespread issue of cross-reactivity and insufficient validation. Through this process, we identified and validated a highly specific antibody for HK3, demonstrating its reliability in western blot and immunohistochemistry applications. Using this validated tool, we reveal the distinct localization of HK3 in myeloid cell populations, providing new insights into its potential functional roles in these cells. This work addresses a critical gap in antibody specificity and establishes HK3 as a uniquely expressed gene in myeloid and immune cells and is absent in other cell types under basal conditions. Providing a foundation for future investigations into its context-dependent functions.</p>\",\"PeriodicalId\":50487,\"journal\":{\"name\":\"European Journal of Histochemistry\",\"volume\":\"69 1\",\"pages\":\"\"},\"PeriodicalIF\":2.1000,\"publicationDate\":\"2025-01-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11956552/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"European Journal of Histochemistry\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.4081/ejh.2025.4175\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/3/12 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q4\",\"JCRName\":\"CELL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"European Journal of Histochemistry","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.4081/ejh.2025.4175","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/3/12 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

准确区分具有高序列同一性的同源蛋白仍然是生物医学研究中的一个重大挑战,因为传统抗体往往缺乏足够的特异性,导致潜在的误解。这个问题在己糖激酶的研究中尤为明显,己糖激酶是一类同工酶,通过磷酸化葡萄糖来催化糖酵解的第一步。除了其典型的代谢作用,己糖激酶发挥关键的非糖酵解功能,特别是在癌症生物学。然而,它们独特的组织分布和上下文依赖的作用往往被商业上可获得的抗体的重叠特异性所掩盖,这可能产生误导性的结果。在这项研究中,我们严格评估了一组针对己糖激酶同工酶3 (HK3)的抗体,强调了交叉反应性和不充分验证的普遍问题。通过这个过程,我们鉴定并验证了一种高度特异性的HK3抗体,证明了它在免疫印迹和免疫组织化学应用中的可靠性。利用这个经过验证的工具,我们揭示了HK3在骨髓细胞群体中的独特定位,为其在这些细胞中的潜在功能作用提供了新的见解。这项工作解决了抗体特异性的一个关键缺口,并确立了HK3是髓细胞和免疫细胞中唯一表达的基因,而在基础条件下其他细胞类型中不存在。为将来研究其依赖于上下文的功能提供基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Reliable hexokinase 3 protein detection in human cell lines and primary tissue.

Accurate differentiation of homologous proteins that share high sequence identity remains a significant challenge in biomedical research, as conventional antibodies often lack sufficient specificity, leading to potential misinterpretations. This issue is particularly evident in the study of hexokinases, a family of isoenzymes that catalyze the first step of glycolysis by phosphorylating glucose. Beyond their canonical metabolic roles, hexokinases play critical non-glycolytic functions, especially in cancer biology. However, their unique tissue distributions and context-dependent roles are often obscured by the overlapping specificities of commercially available antibodies, which can produce misleading results. In this study, we rigorously evaluated a panel of antibodies targeting hexokinase isoenzyme 3 (HK3), highlighting the widespread issue of cross-reactivity and insufficient validation. Through this process, we identified and validated a highly specific antibody for HK3, demonstrating its reliability in western blot and immunohistochemistry applications. Using this validated tool, we reveal the distinct localization of HK3 in myeloid cell populations, providing new insights into its potential functional roles in these cells. This work addresses a critical gap in antibody specificity and establishes HK3 as a uniquely expressed gene in myeloid and immune cells and is absent in other cell types under basal conditions. Providing a foundation for future investigations into its context-dependent functions.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
European Journal of Histochemistry
European Journal of Histochemistry 生物-细胞生物学
CiteScore
3.70
自引率
5.00%
发文量
47
审稿时长
3 months
期刊介绍: The Journal publishes original papers concerning investigations by histochemical and immunohistochemical methods, and performed with the aid of light, super-resolution and electron microscopy, cytometry and imaging techniques. Coverage extends to: functional cell and tissue biology in animals and plants; cell differentiation and death; cell-cell interaction and molecular trafficking; biology of cell development and senescence; nerve and muscle cell biology; cellular basis of diseases. The histochemical approach is nowadays essentially aimed at locating molecules in the very place where they exert their biological roles, and at describing dynamically specific chemical activities in living cells. Basic research on cell functional organization is essential for understanding the mechanisms underlying major biological processes such as differentiation, the control of tissue homeostasis, and the regulation of normal and tumor cell growth. Even more than in the past, the European Journal of Histochemistry, as a journal of functional cytology, represents the venue where cell scientists may present and discuss their original results, technical improvements and theories.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信