BrdU細(xì)胞增殖活性檢測方法 |西美杰Exalpha 代理
一?細(xì)胞增殖檢測的意義
細(xì)胞增殖是生物體的重要生命特征，是生物體生長、發(fā)育、繁殖以及遺傳的基礎(chǔ)。監(jiān)測細(xì)胞群的生長對細(xì)胞的狀態(tài)研究很有必要?？蒲泄ぷ髡邆兘?jīng)常需要檢測細(xì)胞增殖情況，比如驗(yàn)證抗腫瘤藥物的藥效時(shí)，監(jiān)測細(xì)胞增殖及存活情況；研究腫瘤侵襲與轉(zhuǎn)移模型時(shí)，分析細(xì)胞轉(zhuǎn)移或侵襲的路徑；分析創(chuàng)傷的修復(fù)或組織的重建時(shí)，檢測細(xì)胞生長情況等等。檢測細(xì)胞增殖及活性變化則可以很好的反映細(xì)胞的生長狀況。
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二 細(xì)胞增殖的研究方法
細(xì)胞增殖的研究方法有很多種，根據(jù)檢測原理不同，主要分為DNA合成檢測法、代謝活性檢測法、細(xì)胞增殖標(biāo)志物檢測法等。本文重點(diǎn)介紹DNA合成檢測之Brdu檢測法，傳統(tǒng)DNA合成檢測方法使用胸腺嘧啶核苷（3H-TdR）摻入DNA，然后通過放射自顯影或閃爍計(jì)數(shù)對DNA進(jìn)行定量，該方法由于存在放射性物質(zhì)、實(shí)驗(yàn)周期長、操作繁瑣，且需要昂貴的設(shè)備，目前已被BrdU（5-Bromo-2-deoxyuridine，5-溴脫氧尿嘧啶核苷）檢測方法取代，BrdU作為一種胸苷類似物，可選擇性地?fù)饺朐鲋臣?xì)胞的DNA中，而且可穩(wěn)定存在，并帶到子代細(xì)胞中，細(xì)胞經(jīng)過固定和變性處理后，可用免疫學(xué)方法檢測DNA中BrdU的含量，從而判斷細(xì)胞的增殖能力。
小編整理匯總了常見細(xì)胞增殖檢測方法，詳見下表。
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三?Exalpha Biologicals |?BrdU檢測試劑盒
因組織細(xì)胞內(nèi)無內(nèi)源性BrdU存在，可利用BrdU相關(guān)抗體來檢測和定位摻入活躍增殖細(xì)胞新合成DNA的BrdU，西美杰代理Exalpha Biologicals?品牌BrdU檢測試劑盒的優(yōu)勢在于使用細(xì)胞層作為固相，將BrdU試劑摻入微孔板中的細(xì)胞，除了對細(xì)胞增殖情況進(jìn)行評估外，還可以從單個(gè)培養(yǎng)物中分析細(xì)胞數(shù)量、形態(tài)和細(xì)胞抗原等信息。同時(shí)Exalpha Biologicals?品牌BrdU檢測試劑盒可通過試劑盒自帶的固定/變性溶液一步完成細(xì)胞固定、透化和DNA變性等實(shí)驗(yàn)流程，大大縮短了實(shí)驗(yàn)時(shí)間。Exalpha Biologicals?品牌BrdU檢測試劑盒中包含檢測細(xì)胞增殖活性所需的所有組分、操作簡便、無放射性、廣泛的樣本適用性，已得到眾多科研工作者的青睞。
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四?Exalpha?Biologicals品牌BrdU檢測試劑盒試劑盒優(yōu)勢
簡便：提供預(yù)稀釋的滴管瓶，現(xiàn)用現(xiàn)配
安全：無放射性成分
完整：包含緩沖液、酶、陽性對照玻片和染色參考玻片
靈活：可用于任何物種組織樣本
靈敏：極低的背景干擾
經(jīng)濟(jì)：成本低，高通量，多種規(guī)格
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經(jīng)BrdU處理的小鼠小腸免疫組織化學(xué)染色圖
西美杰代理的Exalpha?Biologicals成立于1998年，是一家位于美國波士頓的生命科學(xué)公司，專門提供尖端抗體、試劑、試劑盒和定制IgY服務(wù)等。2016年5月Exalpha?Biologicals與Nordic-MUbio達(dá)成長期銷售協(xié)議，Exalpha?Biologicals產(chǎn)品可通過Nordic-MUbio直接銷售給世界各地的客戶。Nordic-MUbio是一家總部位于荷蘭發(fā)展迅猛的抗體試劑公司，Nordic-MUbio提供單克隆抗體、生物標(biāo)志物、即用型流式細(xì)胞術(shù)抗體等產(chǎn)品組合。2022年4月Absolute Biotech將包括Absolute?antibody、LSBio、Kerafast、Exalpha、Nordic-MUbio和Everest在內(nèi)多個(gè)生命科學(xué)品牌聯(lián)合成一個(gè)新公司，為世界各地的科研人員提供獨(dú)特抗體、細(xì)胞系、其他試劑及產(chǎn)品定制服務(wù)。
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參考文獻(xiàn)
Nakamura, S., et al. Application of bromodeoxyuridine (BrdU) and anti-BrdU monoclonal antibody for the in vivo analysis of proliferative characteristics of human leukemia cells in bone marrows. Oncology 1991, 48, 285-289
Wilson, G., Cell kinetic studies using a monoclonal antibody to bromodeoxyuridine. Methods Mol. Biol. 1998, 80, 255-266
Gray, J. (Ed), Special Issue: Monoclonal antibodies against bromodeoxyuridine Cytometry 1985, Vol. 6(6)
Gutschalk, C.M., et al. ‘Granulocyte Colony-Stimulating Factor and Granulocyte-Macrophage Colony-Stimulating Factor Promote Malignant Growth of Cells from Head and Neck Squamous Cell Carcinomas In vivo.’ Cancer Res., 66, 8026-8036 (2006).
Gray, J. (Ed), Special Issue: Monoclonal antibodies against bromodeoxyuridine Cytometry 1985, Vol. 6(6)
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Hawker JR Jr., 'Chemiluminescence-based BrdU ELISA to measure DNA synthesis.' J Immunol Methods. 2003 Mar 1;274(1-2):77-82.
Ang, L.P.K., et al. 'Development of a conjunctival epithelial equivalent with improved proliferative properties using a multistep serum-free culture system.' Investigative Ophthalmology & Visual Science, 2004, 45, 1789-1795
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Wen-Ning Zhao, Norma K. Hylton, Jennifer Wang, Peter S. Chindavong, Begum Alural, Iren Kurtser, Aravind Subramanian, Ralph Mazitschek, Roy H. Perlis, Stephen J. Haggarty. Activation of WNT and CREB Signaling Pathways in Human Neuronal Cells in Response to the Omega-3 Fatty Acid Docosahexaenoic Acid (DHA). Mol Cell Neurosci. 2019 Sep; 99: 103386.
Eleni Venetsanakos, Ken A. Brameld, Vernon T. Phan, Erik Verner, Timothy D. Owens, Yan Xing, Danny Tam, Jacob LaStant, Kwan Leung, Dane E. Karr, Ronald J. Hill, Mary E. Gerritsen, David M. Goldstein, Jens Oliver Funk, and J. Michael Bradshaw. The Irreversible Covalent Fibroblast Growth Factor Receptor Inhibitor PRN1371 Exhibits Sustained Inhibition of FGFR after Drug Clearance Mol Cancer Ther. 2017, 16(12); 2668–76.
Nakamura, S., et al. Application of bromodeoxyuridine (BrdU) and anti-BrdU monoclonal antibody for the in vivo analysis of proliferative characteristics of human leukemia cells in bone marrows. Oncology 1991, 48, 285-289
Wilson, G., Cell kinetic studies using a monoclonal antibody to bromodeoxyuridine. Methods Mol. Biol. 1998, 80, 255-266
Gray, J. (Ed), Special Issue: Monoclonal antibodies against bromodeoxyuridine Cytometry 1985, Vol. 6(6)
Gutschalk, C.M., et al. ‘Granulocyte Colony-Stimulating Factor and Granulocyte-Macrophage Colony-Stimulating Factor Promote Malignant Growth of Cells from Head and Neck Squamous Cell Carcinomas In vivo.’ Cancer Res., 66, 8026-8036 (2006).
Gray, J. (Ed), Special Issue: Monoclonal antibodies against bromodeoxyuridine Cytometry 1985, Vol. 6(6)
