Vari Fluor 633-Phalloidin 产品活性：Vari Fluor 633-Phalloidin 是 Phalloidin 的荧光衍生物，可对肌丝蛋白特异性标记，在标记时可呈现红色荧光 (Ex/Em=630 nm/650 nm)。 产品来源： https://www.medchemexpress.cn/vari-fluor-633-phalloidin.html 研究领域：Others 作用靶点：Fluorescent Dye In Vitro: General Introdction Ghost pen cyclic peptide is a toxin isolated from the deadly umbrella-shaped mushrooms. It is a bicyclic peptide that binds specifically to F-actin. Therefore, the distribution of F-actin can be easily studied by labeling the peptide with a fluorescent dye. Inside the cyclic peptide, an uncommon thioether bridge between cysteine and tryptophan forms an inner ring structure. At elevated pH, the thioether is cleaved and the peptide loses its affinity for actin. Vari Fluor-Phalloidin is a fluorescent derivative of the VF series of dye-labeled phalloidin that specifically labels actin. The fluorescence is highly stable and can be retained in the cell for more than a week. One unit (T) of VF-Phalloidin refers to the amount of dye used to stain a slide loaded with cells. General Protocol 1.Preparation of VF-Phalloidin Working Solution 1.1 Preparation of storage solution Take appropriate amount of methanol or sterile water to dissolve the lyophilized powder in the brown tube to configure the mother liquor. Add 0.25 mL of dissolution solution to 50T dye. Add 1.5 mL of solvent to 300T size dye. Note: VF-Phalloidin storage solution is recommended to be stored at -20 ℃ or -80 ℃ after dispensing and protected from light, and methanol is recommended for long-term storage. 1.2 Preparation of working solution Dilute the stock solution with pre-warmed serum-free cell culture medium or PBS at 1:40-1:200, i.e., dilute 1-5 μL of VF-Phalloidin stock solution per 200 μL of PBS. Note: Please adjust the concentration of VF-Phalloidin working solution according to the actual situation and use it as it is. 2. Cell staining (adherent cells) 2.1 Culture adherent cells on sterile coverslips and wash them 3 times with PBS. 2.2 Fix the cells with PBS solution containing 4% formaldehyde for 20 min at room temperature. Note: Methanol can destroy actin during fixation. Therefore, it is best to avoid fixatives containing any methanol. The preferred fixative is methanol-free formaldehyde. 2.3 Wash the cells 3 times with PBS. 2.4 The cells are permeabilized with 0.4% Triton X-100 in PBS for 10 min at room temperature. 2.5 Wash the cells three times with PBS. 2.6 Add 200 μL of dye working solution, shake gently to completely cover the cells, and incubate at room temperature away from light for 15-45 minutes. Note: The chromosome volume can be adjusted according to the sample. To avoid evaporation of the dye solution during incubation, place the coverslip in a sealed container. 2.7 Pipette off the dye working solution and wash 2-3 times with PBS or culture medium. 2.8 Use fluorescence microscope or flow cytometer to observe under the corresponding channel. Precautions 1.Fluorescently-labeled Ghost Pen Cyclic Peptides are not cell permeable and therefore have not been widely used for live cell labeling. However, it has been reported that living cells may be labeled by cytosolic drinking or by unknown mechanisms. Generally, more dye is required to stain living cells. Alternatively, fluorescently labeled ghost pen cyclic peptides can be injected into cells to monitor actin distribution and cell movement. 2.The actual content of the dye is small, so dissolve it directly in the tube upon receipt and perform the experiment. 3.Please centrifuge the product instantly to the bottom of the tube before use. 4.This product is limited to scientific research use by professionals, and is not to be used for clinical diagnosis or treatment, food or medicine. 5.For your safety and health, please wear lab coat and disposable gloves. 相关产品：MitoSOX Red | BODIPY 493/503 | D-Luciferin potassium | Nile Red | FITC | JC-1 | Propidium Iodide | DiR | FITC-Dextran (MW 4000) | Dihydroethidium | CFDA-SE | 2-NBDG | Calcein-AM | DiI | Thiazolyl Blue | Cy5.5 | TMRE | Fluo-4 AM | CY5 | Indocyanine green | PKH 26 | DiD perchlorate | Acridine Orange hydrochloride | BODIPY 581/591 C11 | FM4-64 | DiO | CY5-SE 品牌介绍: MCE (MedChemExpress) 拥有数百种全球独家化合物，我们致力于为全球科研客户提供最新最全的高品质小分子活性化合物；10,000 多种高选择性抑制剂、激动剂涉及各热门信号通路及疾病领域；设有专业的实验中心和严格的质控、验证体系；提供 LC/MS、NMR、HPLC、手性分析、元素分析等各项质检报告，确保产品的高纯度、高品质；产品的生物活性多经各国客户实验验证；Nature, Cell, Science 等多种顶级期刊及制药专利收录了MCE客户的科研成果；专业团队跟踪最新的制药及生命科学研究进展，为您提供全球最新的活性化合物；与世界各大制药公司及知名科研机构建立了长期的合作。