阿拉丁生产的LumiDye™ SMCC Activated PerCP-Cy5.5 (SMCC-PerCP-Cy5.5)，即SMCC活化PerCP-Cy5.5。PerCP-Cy5.5是一种常用流式荧光染料，由PerCP与Cy5.5（合成花青素染料，发射峰~690 nm）通过化学方法偶联而成。SMCC活化PerCP-Cy5.5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PerCP-Cy5.5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。
PerCP-Cy5.5是流式细胞术中常用的串联荧光染料，其吸收主峰位于约488 nm，发射峰约690 nm，PerCP（多甲藻素-叶绿素-蛋白质复合物）提取自甲藻纲物种，其串联偶联物（如PerCP-Cy5.5）可通过标准488 nm激光激发，并发射远红光波长，适用于细胞多色流式分析。这种多波长发射特性使PerCP-Cy5.5偶联物能与FITC、PE及其他荧光染料协同进行多色分析。
本产品为PerCP-Cy5.5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-PerCP-Cy5.5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-PerCP-Cy5.5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-PerCP-Cy5.5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。
本产品规格为0.1 mg和1 mg，最佳使用浓度为5 mg/mL(加纯水复溶即可)，最佳标记质量比为IgG:SMCC-PerCP-Cy5.5=1:1。注：本产品需现用现配。
图1 LumiDye™ SMCC Activated PerCP-Cy5.5 (SMCC-PerCP-Cy5.5)（M1491734）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. 在正式实验前，需将试剂恢复至室温，较低的温度会影响偶联效果。开启瓶盖之前，先适当离心，将粉末聚集于管底，避免开盖时损失。
5. 操作过程中，务必穿戴实验服、一次性手套及防护装备；
6. 所有产品仅限于研究用途。
Introduction
LumiDye™ SMCC Activated PerCP-Cy5.5 (SMCC-PerCP-Cy5.5) is formed by modifying PerCP-Cy5.5 with Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC). PerCP-Cy5.5 is a commonly used fluorescent dye for flow cytometry, chemically conjugated from PerCP and Cy5.5 (a synthetic cyanin dye with an emission peak at ~690 nm). SMCC Activated PerCP-Cy5.5 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.
PerCP-Cy5.5 is a popular tandem color used in flow cytometry. Its primary absorption peak is at ~488 nm with emission peak at ~690 nm. PerCP (Peridinin-chlorophyll-protein complex) is isolated from Dinophyceae sp. Its tandem conjugates (such as PerCP-Cy5.5) can be excited with a standard 488 nm laser and emits in the far red at a longer wavelength for multicolor flow cytometric analysis of cells.
This product is a purified conjugate of PerCP-Cy5.5 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 to release free thiol groups. Thiol groups can also be added to primary amine groups of biomacromolecule, such as a protein, through chemical reactions.
Two options are available: 0.1 mg and 1.0 mg. The optimal concentration for this product is 5 mg/mL (reconstitute with ultrapure water). For IgG labeling, the recommended mass ratio is IgG: SMCC-PerCP-Cy5.5 = 1:1. Note: The SMCC-PerCP-Cy5.5 solution must be prepared fresh before each use.
Figure 1. Schematic diagram of the conjugation of LumiDye™ SMCC Activated PerCP-Cy5.5 (SMCC-PerCP-Cy5.5) (M1491734）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-PerCP-Cy5.5 should be stored at 4℃ in the dark. Do not freeze.
4. Before formal experimentation, allow all reagents to reach room temperature, as lower temperatures may impair conjugation efficiency. Before opening the bottle cap, centrifuge it briefly to settle the powder at the bottom of the tube, preventing loss during opening.
5. During operation, always wear a lab coat, disposable gloves, and protective equipment.
6. All products are for research use only.
危险品化学品经营许可证（带存储）