Alexa Fluor™750 NHS 酯（琥珀酰亚胺酯）
Alexa Fluor™750 NHS 酯（琥珀酰亚胺酯）
引用和文献 (13)
Invitrogen™

Alexa Fluor™750 NHS 酯（琥珀酰亚胺酯）

Alexa Fluor™ 750 是一种明亮且具有光稳定性的近红外染料，其光谱与 Cy7 染料相似。Alexa Fluor™ 750 染料用于成像和流式细胞分析中稳定信号的生成，具有水溶性和了解更多信息
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货号数量
A201115 mg
A375753 x 100μg
A200111 mg
A3756825 mg
货号 A20111
价格（CNY)
24,578.00
Each
添加至购物车
数量:
5 mg
价格（CNY)
24,578.00
Each
添加至购物车
Alexa Fluor™ 750 是一种明亮且具有光稳定性的近红外染料，其光谱与 Cy7 染料相似。Alexa Fluor™ 750 染料用于成像和流式细胞分析中稳定信号的生成，具有水溶性和 pH 值不敏感性（pH 值 4 - pH 值 10）。该长波长 Alexa Fluor™ 染料的荧光人眼不可见，但很容易被大多数成像系统检测到。除反应性染料制剂外，我们还提供可与多种抗体、肽、蛋白、示踪剂和扩增底物偶联并且针对细胞标记和检测进行优化的 Alexa Fluor™ 750 染料（了解更多信息）。

Alexa Fluor™ 750 的 NHS 酯（或琥珀酰亚胺酯）是将该染料与蛋白或抗体偶联的较常用工具。NHS 酯可用于标记蛋白、胺修饰的寡核苷酸和其他含胺分子的伯胺 (R-NH2)。所得 Alexa Fluor™ 偶联物将显示出比其他光谱相似荧光基团的偶联物更亮的荧光和更高的光稳定性。

关于该 AlexaFluor™ NHS 酯的详细信息：

荧光基团标记：Alexa Fluor™ 750 染料
反应性基团：NHS 酯
反应性：蛋白和配体、胺修饰的寡核苷酸上的伯胺
偶联物的 Ex/Em：753/782 nm
消光系数：290,000 cm-1M-1
光谱类似染料：Cy7
分子量：∼1300

典型偶联反应
您可以将胺反应性试剂与几乎任何蛋白或肽偶联（提供的方案针对 IgG 抗体进行了优化）。您可以针对任何量的蛋白按比例缩放反应，但为了获得最佳结果，蛋白的浓度应至少为 2 mg/mL。我们建议使用三种不同的反应性试剂/蛋白摩尔比进行三种不同程度的标记。

Alexa Fluor™ NHS 酯通常溶于高质量的无水二甲基甲酰胺 (DMF) 或二甲亚砜 (DMSO) (D12345) 中，并在 0.1–0.2 M 碳酸氢钠缓冲液（pH 值 8.3）中于室温下进行反应，持续 1 小时。由于末端胺的 pKa 低于赖氨酸 ε-氨基基团的 pKa，您可以使用接近中性 pH 值的缓冲液对胺末端进行更具选择性的标记。

偶联物纯化
通常使用凝胶过滤柱（如 Sephadex™ G-25、BioGel™ P-30 或等效柱）将标记抗体与游离 Alexa Fluor™ 染料分离。对于更大或更小的蛋白，选择具有适当分子量滤除点的凝胶过滤介质或通过透析纯化。我们提供了多种优化的纯化试剂盒，可用于不同量抗体偶联物：
0.5-1 mg 用抗体偶联物纯化试剂盒 (A33086)
20-50 µg 用抗体偶联物纯化试剂盒 (A33087)
50-100 µg 用抗体偶联物纯化试剂盒 (A33088)

了解关于蛋白和抗体标记的更多信息
我们提供一系列 Molecular Probes™ 抗体和蛋白标记试剂盒，旨在满足您的起始材料和实验设置需求。参见我们的抗体标记试剂盒或使用我们的标记化学选择工具进行其他选择。欲了解有关我们标记试剂盒的更多信息，请参阅 Molecular Probes™ 手册中第 1.2 节—蛋白和核酸标记试剂盒。

我们还’可为您定制偶联物
如果您’无法在我们的在线目录中找到’想要的产品，我们还’可为您定制抗体或蛋白偶联物。我们的定制偶联服务是高效和保密的，我们保证我们的工作质量。我们经过ISO 9001:2000认证。
仅供科研使用。不可用于诊断程序。
规格
化学反应性胺
发射782 nm
激发753 nm
标签或染料Alexa Fluor™ 750
产品类型染料
数量5 mg
反应一部分活性酯、琥珀酰亚胺酯
运输条件室温
标签类型Alexa Fluor 染料
产品线Alexa Fluor
Unit SizeEach
内容与储存
储存在冰箱(-5 至 -30°C)中并避光。

常见问题解答 (FAQ)

I am labeling a protein with Alexa Fluor 488 SDP ester. The manual recommends using a sodium bicarbonate buffer at pH 8.3. Can I use a different buffer instead?

Yes. The important thing is to use a buffered solution with a pH between 8.0 and 8.5. Do not use Tris buffer, which has amine groups. Most other buffers will work fine in that pH range. This is also true for other amine-reactive dyes, such as succinimidyl (NHS) esters or TFP esters.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am not going to use all of my Alexa Fluor succinimidyl ester reactive dye. Can I just make it up in DMSO and store aliquots at -20 degrees C?

This is not recommended. Any trace amounts of water in the DMSO can promote spontaneous hydrolysis over time. Even if using anhydrous DMSO, DMSO is hygroscopic; it readily absorbs moisture from the atmosphere over time. A better alternative is to dissolve the reactive dye in a volatile solvent, make smaller aliquots and then evaporate off the solvent using a vacuum pump. The smaller aliquots of solid reactive dye should then be stored frozen, desiccated and protected from light. Contact Technical Support by sending an email to techsupport@thermofisher.com for the recommended volatile solvent.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

引用和文献 (13)

引用和文献
Abstract
Tyramide signal amplification for analysis of kinase activity by intracellular flow cytometry.
Authors:Clutter MR, Heffner GC, Krutzik PO, Sachen KL, Nolan GP,
Journal:Cytometry A
PubMed ID:20824632
'Intracellular flow cytometry permits quantitation of diverse molecular targets at the single-cell level. However, limitations in detection sensitivity inherently restrict the method, sometimes resulting in the inability to measure proteins of very low abundance or to differentiate cells expressing subtly different protein concentrations. To improve these measurements, an enzymatic amplification ... More
Quantitative comparison of long-wavelength Alexa Fluor dyes to Cy dyes: fluorescence of the dyes and their bioconjugates.
Authors:Berlier JE, Rothe A, Buller G, Bradford J, Gray DR, Filanoski BJ, Telford WG, Yue S, Liu J, Cheung CY, Chang W, Hirsch JD, Beechem JM, Haugland RP, Haugland RP
Journal:J Histochem Cytochem
PubMed ID:14623938
'Amine-reactive N-hydroxysuccinimidyl esters of Alexa Fluor fluorescent dyes with principal absorption maxima at about 555 nm, 633 nm, 647 nm, 660 nm, 680 nm, 700 nm, and 750 nm were conjugated to antibodies and other selected proteins. These conjugates were compared with spectrally similar protein conjugates of the Cy3, Cy5, ... More
Fluorescent cell barcoding in flow cytometry allows high-throughput drug screening and signaling profiling.
Authors:Krutzik PO, Nolan GP
Journal:Nat Methods
PubMed ID:16628206
'Flow cytometry allows high-content, multiparameter analysis of single cells, making it a promising tool for drug discovery and profiling of intracellular signaling. To add high-throughput capacity to flow cytometry, we developed a cell-based multiplexing technique called fluorescent cell barcoding (FCB). In FCB, each sample is labeled with a different signature, ... More
Noninvasive tracking of donor cell homing by near-infrared fluorescence imaging shortly after bone marrow transplantation.
Authors:Ushiki T, Kizaka-Kondoh S, Ashihara E, Tanaka S, Masuko M, Hirai H, Kimura S, Aizawa Y, Maekawa T, Hiraoka M,
Journal:PLoS One
PubMed ID:20559437
'BACKGROUND: Many diseases associated with bone marrow transplantation (BMT) are caused by transplanted hematopoietic cells, and the onset of these diseases occurs after homing of donor cells in the initial phase after BMT. Noninvasive observation of donor cell homing shortly after transplantation is potentially valuable for improving therapeutic outcomes of ... More
A Small Molecule Inhibitor of Polycomb Repressive Complex 1 Inhibits Ubiquitin Signaling at DNA Double-strand Breaks.
Authors:Ismail IH, McDonald D, Strickfaden H, Xu Z, Hendzel MJ,
Journal:
PubMed ID:23902761
'Polycomb-repressive complex 1 (PRC1)-mediated histone ubiquitylation plays an important role in aberrant gene silencing in human cancers and is a potential target for cancer therapy. Here we show that 2-pyridine-3-yl-methylene-indan-1,3-dione (PRT4165) is a potent inhibitor of PRC1-mediated H2A ubiquitylation in vivo and in vitro. The drug also inhibits the accumulation ... More
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