Vari Fluor 565 SE 产品活性：Vari Fluor 565 SE (VF 565 SE) 是 Vari Fluor SE 系列的标记染料 (Ex/Em=563 nm/594 nm)。Vari Fluor SE 系列染料是一类含有 NHS 酯基的荧光染料，用于标记抗体、蛋白质、肽、胺修饰的寡核苷酸和其他生物分子上的游离胺 (-NHX)。 产品来源： https://www.medchemexpress.cn/vari-fluor-565-se.html 研究领域：Others 作用靶点：Fluorescent Dye In Vitro: Protocol 1.Protein Preparetion 1) In order to obtain the best labeling effect, please prepare the protein (antibody) concentration as 2mg/mL. 2) The pH value of protein solution shall be 8.5±0.5. If the pH is lower than 8.0, 1 M sodium bicarbonate shall be used for adjustment. 3) If the protein concentration is lower than 2 mg/mL, the labeling efficiency will be greatly reduced. In order to obtain the best labeling efficiency, it is recommended that the final protein concentration range is 2-10 mg/mL. 4) The protein must be in the buffer without primary amine (such as Tris or glycine) and ammonium ion, otherwise the labeling efficiency will be affected. 2.Dye Preparation（Example for VF 488 SE） Add anhydrous DMSO into the vial of VF 488 SE to make a 10 mg/mL stock solution. Mix well by pipetting or vortex. 3.Calculation of dye dosage The amount of VF 488 SE required for reaction depends on the amount of protein to be labeled, and the optimal molar ratio of VF 488 SE to protein is about 10. Example: assuming the required marker protein is 500 μL 2 mg/mL IgG (MW=150,000), use 100 μL DMSO dissolve 1 mg VF 488 SE, the required VF 488 SE volume is 5.05 μL, and the detailed calculation process is as follows: 1) mmol (IgG) = mg/mL (IgG)×mL (IgG) / MW (IgG) =2 mg/mL × 0.5 mL / 150,000 mg/mmol= 6.7×10-6 mmol 2) mmol (VF 488 SE) = mmol (IgG)× 10 = 6.7×10-6 mmol×10 = 6.7 × 10-5 mmol 3) μL (VF 488 SE) = mmol (VF 488 SE)×MW (VF 488 SE) / mg/μL (VF 488 SE) = 6.7 ×10-5 mmol ×834 mg/mmol / 0.01 mg/μL = 5.6 μL (VF 488 SE) 4.Run conjugation reaction 1) A good volume of freshly prepared 10 mg/mL VF 488 SE is slowly added to 0.5 mL protein sample. In solution, gently shake to mix, then centrifuge briefly to collect the sample at the bottom of the reaction tube. Don't mix well to prevent protein samples from denaturation and inactivation. 2) The reaction tubules were placed in a dark place and incubated gently at room temperature for 60 minutes at intervals.For 10-15 minutes, gently reverse the reaction tubules several times to fully mix the two reactants and raise the bar efficiency. 5.Purify the conjugation The following protocol is an example of dye-protein conjugate purification by using a Sephadex G-25 column. 1）Prepare Sephadex G-25 column according to the manufacture instruction. 2）Load the reaction mixture (From 'Run conjugation reaction') to the top of the Sephadex G-25 column. 3）Add PBS (pH 7.2-7.4) as soon as the sample runs just below the top resin surface. 4）Add more PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate. Note 1.CY dyes is sensitive to light and humidity. Immediately add CY dyesr solution and discard the unused part. 2.Low concentrations of sodium azide (≤3 mM or 0.02%) or thiomersal (≤0.02 mM or 0.01%) did not significantly interfere with protein labeling; However, 20-50% glycerol will reduce labeling efficiency. 3.Avoid buffering with primary amines (e.g., Tris, glycine) or ammonium ions,It compete with labeled proteins. 4.This product is only for scientific research by professionals, and shall not be used in 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。