Biotium/AccuClear Ultra High Sensitivity dsDNA Quantitation Kit with 1 DNA Standard/31029/2000-assay

来自 : 发布时间：2025-01-16

返回突出显示的引文组织切片中红细胞自发荧光的有效淬灭相关页面CF® 染料下一代荧光染料跨越可见光和近红外光谱一抗和二抗偶联物越来越多的数千种标记一抗和二抗的集合EverBrite™ 封片剂技术提示：对抗组织自发荧光SHARE ARTICLEShare这篇文章与一位同事。随着新型染料和成像技术的发展，荧光显微术在研究和临床诊断中变得越来越重要。然而，自发荧光仍然是一个潜在的问题，特别是在依赖固定哺乳动物组织的研究中。它表现为强背景荧光，跨越多个通道，会干扰几种常用荧光报告基因的信号，并使数据分析复杂化。细胞外基质成分、红细胞 (RBC)、脂褐素、色素细胞和纤维蛋白xation 方法都可以促进自发荧光。已经开发了几种淬灭剂和组织处理方法作为对策，但收效有限。组织或整个生物体灌注以冲洗掉干扰性红细胞对胚胎组织或临床样本的使用有限。在神经科学当前协议的最新报告中，作者展示了 TrueBlack® 如何在固定的胚胎组织中有效地淬灭跨多个波长的红细胞自发荧光, 不会干扰免疫荧光信号强度或引入背景染色。他们还证明 TrueBlack® 与多色免疫荧光染色兼容，而不是苏丹黑等常用猝灭剂，并提供有关常规免疫荧光显微镜的最佳浓度、孵育时间和多种使用条件的详细信息。要阅读原始文章，请单击此处.Whittington, N. C., & Wray, S. (2017)。抑制红色用于固定小鼠胚胎组织免疫细胞化学的血细胞自发荧光。当前的神经科学协议，81,2.28.1–2.28.12。 doi: 10.1002/cpns.35 在用兔抗 GFAP 染色的人大脑皮层的甲醇固定冷冻切片进行免疫荧光染色后，使用 TrueBlack® 后处理淬灭脂褐质自发荧光，然后用 CF®640R 山羊抗兔抗体和 DAPI 安装在 EverBrite™ 封固剂中。了解有关 Biotium 的 TrueBlack® 脂褐素自发荧光猝灭剂的更多信息。另请参阅我们的技术提示：对抗组织自发荧光。返回重点引文用于近红外小动物成像的便携式基于 LED 的光声成像系统相关页面DiRCellBrite® NIR 细胞质膜染料Annexin V 近红外 CF® 染料偶联物近红外 CF® 染料游离酸VivoBrite™ 用于小动物体内成像的快速抗体标记试剂盒DCDAPHSHARE ARTICLEShare复制链接发送电子邮件光声成像 (PAI) 是一种非侵入性、非电离、高分辨率成像技术，它结合了光吸收/激发（来自血红蛋白或黑色素等内源性发色团，或外源性造影剂）与超声检测。由于超声波的散射较低，PAI 可以提供高分辨率的光学对比成像，显示出作为 MRI 和 CT 扫描替代品的真正前景。随着分子成像、肿瘤学、眼科、心脏病学和神经学领域的应用不断增长，最近的一份出版物描述了一种基于 LED 的便携式光声用于商业应用的抽动成像系统。通过用多波长 LED 代替激光器作为光激发源，作者能够使成像系统紧凑、便携且价格合理，可广泛用于临床。体内视网膜血管监测，以及用近红外造影剂 (DiR) 标记的人间充质干细胞成像在活小鼠身上得到证实。要阅读原文，请单击此处。Hariri A、Lemaster J、Wang J、 Jeevarathinam AS、Chao DL、Jokerst JV。一种经济且便携的基于 LED 的光声成像系统的表征，以促进分子成像。光声学。 2017 年 11 月 26 日；9:10-20。 doi:10.1016/j.pacs.2017.11.001.HeLa 细胞用 Biotium 的近红外细胞质膜染料 CellBrite™ NIR680 染色。使用 Hoechst 33342 对细胞核进行复染。图像是在 Zeiss LSM 700 共聚焦显微镜上采集的。详细了解用于体内成像的 Biotium 近红外染料和 CF® 染料偶联物。返回突出显示的引文 DAPI 和 Hoechst 染料的 UV 诱导光转化伪影会导致绿色和红色荧光发射colleague.Copy LinkSend EmailDAPI 和 Hoechst 染料在荧光显微镜中通常用作染色体或核染色剂。它们具有相似的激发和发射光谱，并对 DNA 表现出高度特异性。通常，在与 DNA 结合时，这些染料在紫外线范围内被最大激发，并在蓝色范围内发射。据报道，紫外线照射可以将这些染料转化为被蓝光激发并发出绿色荧光的形式，从而导致成像组合/多色实验中的伪影和可能混淆的数据解释。最近的一项研究报告说，这些染料也可以转化为形式 t帽子可以被绿光激发并发出红色荧光。DAPI 在暴露于紫外光后光转化为绿色和红色荧光。在通过 DAPI 滤光片立方体暴露于汞灯激发 3 分钟之前或之后，用 DAPI 染色的固定和透化 HeLa 细胞通过共聚焦显微镜成像。为了最大限度地减少与这些染料相关的潜在光转换伪影，请参阅我们的技术提示避免 DAPI 的 UV 光转换产生的伪影和 Hoechst。要阅读原文，请单击此处。Karg, T.J. & Golic, K.G. Chromosoma (2017) DAPI 和 Hoechst 染料在暴露于紫外线激发后光转化为绿色和红色发光形式。 https://doi.org/10.1007/s00412-017-0654-5 详细了解 Biotium 的商标和经典核染色剂。AccuClear™ Ultra High Sensitivity dsDNA Quantitation Solution utilizes a novel, next-generation DNA binding dye for highly sensitive and accurate quantitation of purified dsDNA samples across a broad range of DNA concentrations. This assay has a linear range between 0.03 ng and 250 ng of dsDNA per assay in microplate format, making it the most versatile DNA quantitation assay.

AccuClear™ Ultra High Sensitivity dsDNA Quantitation Solution utilizes a novel, next-generation DNA binding dye for highly sensitive and accurate quantitation of purified dsDNA samples across a broad range of DNA concentrations. This assay has a linear range between 0.03 ng and 250 ng of dsDNA per assay in microplate format, making it the most versatile DNA quantitation assay. Unlike absorbance-based measurements, AccuClear™ dye is highly selective for double-stranded DNA over single stranded DNA or RNA.

The AccuClear™ dsDNA quantitation assay is designed for use with fluorescence 96-well plate readers equipped with excitation and emission filters for detecting green fluorescence. The unique spectral properties of AccuClear™ dye make it especially well-suited for use with instruments with blue LED excitation sources. AccuClear™ also is compatible with handheld fluorometers such as Invitrogen’s Qubit® and Promega’s QuantiFluor®-P, however the standard curve calibration programs for these instruments may not cover the full dynamic range of the AccuClear™ kit standard curve. For DNA quantitation using the Qubit® we recommend our AccuGreen™ High Sensitivity dsDNA Quantitation Kit (catalog no. 31066), which was formulated specifically for that instrument.

The AccuClear™ Ultra High Sensitivity dsDNA Quantiation Kit with 1 DNA Standard contains AccuClear™ dye, 20X assay buffer, and a 25 ng/uL calf thymus dsDNA standard stock solution that can be used to prepare a set of standards. Biotium also offers the AccuClear™ Ultra High Sensitivity dsDNA Quantiation Kit with 7 DNA Standards (1000 assays, catalog no. 31028), which includes AccuClear™ dye, 1X assay buffer, and a set of prediluted calf thymus dsDNA standards. AccuClear™ Ultra High Sensitivity dsDNA Quantitation Solution (1000 assays, catalog no. 31027), which includes dye and 1X buffer without standards, is available for customers who wish to prepare their own DNA standards.

Components

• AccuClear™ Dye (100X)
• AccuClear™ Buffer (20X)
• Calf Thymus dsDNA Standard, 25 ng/uL

Features

• Linear range: 0.03-250 ng dsDNA
• Green fluorescence (Ex/Em 460/507 nm)
• Unrivaled sensitivity and dynamic range
• Novel green fluorescent dye is a perfect match for blue LED excitation sources
• Green fluorescence detection compatible with fluorescence microplate reader, handheld fluorometer, or NanoDrop® fluorospectrophotometer.

Other DNA Quantitation Kits:

dsDNA Quantitation KitLinear detection range (dsDNA in assay)Ex/Em (nm)Features

AccuBlue™ NextGen	1 pg*-3 ng	468/507	Quantitation of extremely precious or dilute samples. Optimal for sensitive applications such as NGS or digital PCR. *Limit of detection in the range of 1 pg to 5 pg depending on instrument sensitivity.
AccuClear™ Ultra High Sensitivity	30 pg-250 ng	468/507	This kit offers great sensitivity and the widest linear range.
AccuBlue™ High Sensitivity	0.2-100 ng	485/530	Quantitation of dsDNA from 0.2-100 ng, with a safer, membrane impermeable DNA dye.
AccuGreen™ High Sensitivity	0.1-100 ng	502/523	Quantitation of DNA using the Qubit® fluorometer from Thermo Fisher.
AccuBlue™ Broad Range	2-2000 ng	350/460	For quantitation of high concentration DNA in a broad linear range with blue fluorescence. Assay can be extended to 4000 ng with minor loss of linearity.

AccuClear kits are intended for quantitation of purified dsDNA samples. For cell quantitation assays, please see our cell viability and cytotoxicity assays.

AccuClear™ technology is covered by granted U.S. and international patents.

References

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