{"title":"利用人黑色素瘤细胞裂解液建立人酪氨酸酶活性抑制试验。","authors":"Gengxuan Shi, Yunjiang Feng, Kathryn F Tonissen","doi":"10.1080/07366205.2024.2441637","DOIUrl":null,"url":null,"abstract":"<p><p>Tyrosinase (TYR) inhibitors are required to treat skin hyperpigmentation. Currently live cell-based TYR assays and mushroom TYR <i>in vitro</i> assays are the common methods used to screen for TYR inhibitors. However, these methods are either time consuming and expensive or are not human TYR (<i>hs</i>TYR) specific. Here, we describe a simple <i>hs</i>TYR assay using cell lysate prepared from pigmented human melanoma cell lines that takes less than 3 hours to complete after collecting cell pellets. We confirmed the assay is species specific by using a known <i>hs</i>TYR inhibitor, kojic acid, as a positive control, while arbutin, which inhibits mushroom TYR, but not <i>hs</i>TYR, was not effective. This assay is a simple method to confirm <i>hs</i>TYR inhibition before conducting follow-up studies in live biological models.</p>","PeriodicalId":8945,"journal":{"name":"BioTechniques","volume":" ","pages":"547-551"},"PeriodicalIF":2.2000,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a human tyrosinase activity inhibition assay using human melanoma cell lysate.\",\"authors\":\"Gengxuan Shi, Yunjiang Feng, Kathryn F Tonissen\",\"doi\":\"10.1080/07366205.2024.2441637\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Tyrosinase (TYR) inhibitors are required to treat skin hyperpigmentation. Currently live cell-based TYR assays and mushroom TYR <i>in vitro</i> assays are the common methods used to screen for TYR inhibitors. However, these methods are either time consuming and expensive or are not human TYR (<i>hs</i>TYR) specific. Here, we describe a simple <i>hs</i>TYR assay using cell lysate prepared from pigmented human melanoma cell lines that takes less than 3 hours to complete after collecting cell pellets. We confirmed the assay is species specific by using a known <i>hs</i>TYR inhibitor, kojic acid, as a positive control, while arbutin, which inhibits mushroom TYR, but not <i>hs</i>TYR, was not effective. This assay is a simple method to confirm <i>hs</i>TYR inhibition before conducting follow-up studies in live biological models.</p>\",\"PeriodicalId\":8945,\"journal\":{\"name\":\"BioTechniques\",\"volume\":\" \",\"pages\":\"547-551\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2024-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"BioTechniques\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1080/07366205.2024.2441637\",\"RegionNum\":4,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/12/16 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"BioTechniques","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1080/07366205.2024.2441637","RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/12/16 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Development of a human tyrosinase activity inhibition assay using human melanoma cell lysate.
Tyrosinase (TYR) inhibitors are required to treat skin hyperpigmentation. Currently live cell-based TYR assays and mushroom TYR in vitro assays are the common methods used to screen for TYR inhibitors. However, these methods are either time consuming and expensive or are not human TYR (hsTYR) specific. Here, we describe a simple hsTYR assay using cell lysate prepared from pigmented human melanoma cell lines that takes less than 3 hours to complete after collecting cell pellets. We confirmed the assay is species specific by using a known hsTYR inhibitor, kojic acid, as a positive control, while arbutin, which inhibits mushroom TYR, but not hsTYR, was not effective. This assay is a simple method to confirm hsTYR inhibition before conducting follow-up studies in live biological models.
期刊介绍：
BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies.
Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers:
Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions
Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices
Surveys of technical approaches related to broad fields of research
Reviews discussing advancements in techniques and methods related to broad fields of research
Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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