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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
产品介绍
阿拉丁生产的LumiDye™ SMCC Activated PE-Cy7 (SMCC-PE-Cy7)，即SMCC活化PE-Cy7，是由4-(N-马来酰亚胺基甲基)环已烷-1-羟酸琥珀酰亚胺酯(Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate, SMCC)修饰PE-Cy7形成，可与含巯基的分子如蛋白等偶联，偶联物常用于荧光显微检测、荧光免疫测定、双色或多色荧光分析、癌细胞表面抗原检测、流式细胞荧光测定等抗体荧光标记以及活体成像等诊断及生物工程技术。
PE-Cy7是一种远红光发射串联荧光团，由藻红蛋白（PE）与Cy7结合而成。供体分子PE可被488 nm蓝光、532 nm绿光或561 nm黄绿色激光激发，并将能量传递给受体分子Cy7，后者发出的荧光可通过780/60 nm带通滤光片捕获。PE-Cy7的激发峰位于565 nm，发射峰位于774 nm。
本产品为PE-Cy7与SMCC的连接产物，经纯化并去除过量的SMCC，溶于含3%海藻糖的PBS-EDTA中，SMCC-PE-Cy7最终以冻干粉的形式提供，长期储存也不损失巯基反应性，保证了SMCC-PE-Cy7与其它含巯基分子如蛋白质等偶联的最佳性质。
使用方法
SMCC-PE-Cy7具有巯基反应性，可以针对抗体等生物大分子进行偶联，这些分子具有半胱氨酸，可以通过还原剂处理以产生游离的巯基基团。当蛋白等生物大分子具有可用的伯胺基团时，也可以通过化学反应添加所需的巯基基团。
本产品最佳使用浓度为5mg/mL，加入200 µL 纯水，摇晃瓶体使染料完全溶解后可进行标记。注：本产品需现用现配。
图1 LumiDye™ SMCC Activated PE-Cy7 (SMCC-PE-Cy7)（M1491729）与蛋白质的偶联原理图
保存条件
4ºC避光保存，12个月内有效，切勿-20ºC或更低温度保存。
注意事项
1. 避免在反应过程中使用含有伯胺和巯基的缓冲液，以减少不必要的竞争反应；
2. 荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光的淬灭；
3. 本产品的偶联产物，须避光保存，请勿冷冻；
4. 操作过程中，务必穿戴实验服、一次性手套及防护装备；
5. 所有产品仅限于研究用途。
Introduction
LumiDye™ SMCC Activated PE-Cy7 (SMCC-PE-Cy7) is formed by modifying PE-Cy7 with Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC). It contains a maleimide group that reacts with thiol groups of target molecules, such as proteins. This product is commonly used in fluorescence microscopy, fluorescence immunoassays, dual or multicolor fluorescence analysis, cancer cell surface antigen detection, flow cytometry fluorescence assays, antibody fluorescence labeling, as well as diagnostics and biotechnology applications such as live cell imaging.
PE-Cy7 is a far red-emitting tandem fluorophore that combines phycoerythrin (PE) and Cy7. The donor molecule, PE can be excited by the 488-nm blue, 532-nm green, or 561-nm yellow-green laser and and transfers energy to the acceptor molecule, Cy7, which emits light that can be captured with a 780/60 nm bandpass filter. PE-Cy7 has an excitation peak at 565 nm and an emission peak at 774 nm.
This product is a purified conjugate of PE-Cy7 and SMCC. After purification to remove excess SMCC, it is dissolved in PBS-EDTA with 3% trehalose and ultimately supplied as a lyophilized powder. The long-term storage of this product does not affect its thiol reactivity, ensuring the optimal properties for conjugation with other thiol-containing molecules such as proteins.
Instruction for use
This product is chemically activated for easy conjugation to biomacromolecules such as antibodies. These molecules contain cysteine residues. Disulfide bonds formed between cystein residues can be treated with a reducing agent, such as DTT and β-mercaptoethanol, to release free thiol groups. Thiol groups can also be added to primary amine groups of biomacromolecule, such as a protein, through chemical reactions.
The optimal concentration for this product is 5 mg/mL. Add 200 μL of ultrapure water, gently swirl the bottle to ensure complete dissolution of the dye, then proceed with labeling. Note: The SMCC-PE-Cy7 solution must be prepared fresh before each use.
Figure 1. Schematic diagram of the conjugation of LumiDye™ SMCC Activated PE-Cy7 (SMCC-PE-Cy7) (M1491729) to proteins.
Storage
Store at 4℃ in the dark for up to 12 months. Do not store at -20℃ or lower temperatures.
Matters needing attention
1. Avoid using buffers containing primary amines and thiols during the reaction to minimize unnecessary competitive reactions.
2. Fluorescent dyes are prone to quenching. Please store and use them in the dark to slow down fluorescence quenching.
3. Products conjugated with the SMCC-PE-Cy7 should be stored at 4℃ in the dark. Do not freeze.
4. During operation, always wear a lab coat, disposable gloves, and protective equipment.
5. All products are for research use only.
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0231994 |
分析证书 | M1491729 |
| ZJ26F0231995 |
分析证书 | M1491729 |
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