BioReagent 级 提供 ≥80%(HPLC) 纯度，适用于对基线干扰要求严格的色谱和分析工作流程。
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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。
CytoTrace Red CMTPX 化学性质与CellTracker CMTPX相同，可自由穿过细胞膜进入细胞内，并在细胞内转化为不可透过细胞膜的产物。该染料能在活细胞中保留多代，可传递至子代细胞，但不会扩散至相邻细胞。
CytoTrace Red CMTPX 染料的荧光标记可持续至少72小时，具有理想的细胞示踪特性。此外，Red CMTPX染料的激发和发射光谱与绿色荧光蛋白(GFP)光谱能很好地分开，适用于多色荧光标记实验。
实验方案
操作步骤
本方案为通用指导，需根据具体实验需求调整
1.准备2-10 mM DMSO储备溶液
向50 μg试剂瓶中加入36 μL DMSO，配制成2 mM储备液（1 mg/mL≈1.46 mM）
注意：储备液需现配现用，剩余溶液应分装避光保存于<-20°C，避免反复冻融。
2.配制工作液
使用前将储备液用HHBS缓冲液（含20 mM Hepes，pH 7.0）或自选缓冲液稀释至1-20 μM，涡旋混匀。
3.用流式细胞仪或荧光显微镜分析细胞：
3.1用测试化合物处理细胞所需的时间。
3.2离心细胞，每管得到2-10 x 10⁵ 细胞。
3.3用500 μL工作液重悬细胞（来自步骤2）。
3.4将细胞与染料溶液在室温或37°C下避光放置15至30分钟。
3.5从细胞中除去染料工作溶液，用HHBS或您选择的缓冲液洗涤细胞。将细胞重悬于500 µL预热的HHBS或培养基中，每管可得到2-10 x 10⁵ 个细胞。
3.6用流式细胞仪或荧光显微镜监测Ex / Em = 577/602nm处的荧光变化。
细菌细胞染色：
推荐将储备液以1:800比例稀释于经测试菌预培养的营养肉汤中（新鲜肉汤或PBS也可用）。菌悬液需用PBS稀释至10⁵-10⁷个/mL，染色方法：将1 mL菌液过0.45 μm滤膜（25mm）后真空抽滤，加入1 mL染料工作液，室温避光孵育5-10分钟，即可对细菌染色。
CytoTrace Red CMTPX is chemically same to CellTracker CMTPX. CytoTrace Red CMTPX freely passes through cell membranes into cells, where it is transformed into cell-impermeant reaction products. CytoTrace Red CMTPX dye is retained in living cells through several generations. The dye is transferred to daughter cells but not adjacent cells in a population. CytoTrace Red CMTPX dye is designed to display fluorescence for at least 72 hours, and the dye exhibits ideal tracking properties. Additionally, the excitation and emission spectra of Red CMTPX Red CMPTX dye are well separated from GFP (green fluorescent protein) spectra allowing for multiplexing.
Sample Assay Protocol
Note: The following is the recommended protocol. It only provides a guideline, should be modified according to the specific needs.
1. Prepare 2-10 mM DMSO stock solution
Add 36 uL DMSO into a 50 μg vial to make 2 mM stock solution (1 mg/ml is equivalent to 1.46 mM)
Note: The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at ≤ -20 °C. Avoid repeated freeze-thaw cycles, and protect from light.
2. Prepare dye working solutionPrepare a 1 to 20 μM dye working solution right before use by diluting the DMSO stock solution from Step 1 with Hanks and 20 mM Hepes buffer (HHBS) or the buffer of your choice, pH 7. Mix them well by vortexing.
3. Analyze cells with a flow cytometer or a fluorescence microscope:
3.1 Treat cells with test compounds for a desired period of time.
3.2 Centrifuge the cells to get 2-10 × 10⁵ cells per tube.
3.3 Resuspend cells in 500 μL of the dye working solution (from Step 2).
3.4 Incubate cells with a dye solution at room temperature or 37 °C for 15 to 30 min, protected from light.
3.5 Remove the dye working solution from the cells, wash the cells with HHBS or buffer of your choice. Resuspend cells in 500 μL of pre-warmed HHBS or medium to get 2-10 × 10⁵ cells per tube.
3.6 Monitor the fluorescence changes at Ex/Em = 577/602 nm by flow cytometry or fluorescence microscope.
Note: For bacterial cells staining: Staining is most efficient when stock solution is diluted 1:800 in nutrient broth preconditioned by overnight growth with the test bacteria, but fresh nutrient broth or PBS may also be used. Bacterial suspensions should be diluted with PBS to 10⁵ - 10⁷ organisms per ml. Bacteria may be stained by applying one ml of solution to .45 um filter (25mm) and vacuum filtering to remove solution, then adding 1ml of dye solution and incubating 5 - 10 minutes at room temperature.
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