DiI-Ac-LDL 红色荧光标记乙酰化低密度脂蛋白/EPC内皮组细胞标记物

— —清道夫受体/巨噬细胞泡沫细胞形成/内皮细胞/小神经胶质细胞     

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想了解内皮祖细胞（EPC）鉴定的双标记方法，可见MKbio整理的产品专题：EPC双标记鉴定产品专题。

搜索关键词：

HDL；LDL；DiI-LDL；氧化修饰 Ox-LDL；乙酰化修饰Ac-LDL；Scavenger receptor 清道夫受体；Cholesterol 胆固醇；Cholesteryl ester (CE) 胆固醇酯；Triglyceride (TG)甘油三酯；Cardiovascular diseases心血管疾病；Atherosclerosis (AS) 动脉粥样硬化；Macrophages 巨噬细胞；FITC-UEA-I 绿色荧光标记荆豆凝集素；

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温馨提示：由于脂蛋白的保存周期比较短，约6周。本司一般都是接到订单后再生产，从而保证客户拿到最长效期的产品。所以，我们货期至少要2天以上。

基本描述：

脂蛋白（Lipoproteins）是血脂在血液中存在、转运及代谢的形式。血清脂蛋白通过超高速离心，并且根据密度大小差异，将脂蛋白分为乳糜微粒（Chylomicron，CM）、极低密度脂蛋白（VLDL）、低密度脂蛋白（LDL）、高密度脂蛋白（HDL）、以及极低密度脂蛋白的代谢中间产物-中间密度脂蛋白（IDL）。

这些脂蛋白之间蛋白到脂质的含量不同，特别是在载脂蛋白（apoproteins）和脂质含量不同【见总表1-血浆脂蛋白特性】。

DiI-Ac-LDL（红色荧光标记乙酰化修饰低密度脂蛋白）的应用：

乙酰化的LDL（Acetylated LDL，Ac-LDL）是修饰LDL中的一种，LDL载脂蛋白上的赖氨酸残基被乙酰化修饰后，不再能结合到LDL受体，进而被含特异性识别修饰LDL的清道夫受体的巨噬细胞和内皮细胞摄取（消化），导致大量带负电荷的脂质小滴进入，产生类似动脉粥样硬化斑块中发现的泡沫样细胞。

本品是红色荧光探针DiI（1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanineperchlorate）标记的人源乙酰化LDL（Human Ac-LDL）。细胞被DiI-Ac-LDL标记后，溶酶体酶降解脂蛋白，使得探针DiI聚集在细胞内膜上。DiI-Ac-LDL可用来标记巨噬细胞和血管内皮细胞，从而能在混合细胞群中鉴定和分选出这些细胞。也可用来研究不同细胞系对乙酰化LDL的内吞作用。

相关产品：更大包装或产品技术问题，请来电/在线咨询，电话：021-54736159 。网站：www.maokangbio.com。

注意事项

为了您的安全和健康，请穿实验服并戴一次性手套操作。

— —Written/Edited by V. Shallan【版权归MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作，主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本，以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。

文献引用：

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After washing, cells were plated on culture dishes at the density of 5×10 6 and cultured with Microvascular Endothelial Cell Growth Medium-2 (EGM-2 MV) BulletKit (Lonza, Walkersville, MD, USA) in a humidified atmosphere of 5% CO2 at 37°C. Two days later, non-adherent cells were removed by washing with PBS twice and the culture medium was replaced every 3 d. After culturing for 1 d, 7 d, 14 d and 28 d, EPCs were identified by optical 

microscope observation, immunofluorescence for both Dil-Acetylated Low Density Lipoprotein (Dil-Ac- LDL; MaoKang Biotechnology, Shanghai, China) and FITC labeled Ulex Europaeus Agglutinin 1 (FITC-UEA-1; MaoKang Biotechnology, Shanghai, China), and flow cytometry (FACSCalibur Flow Cytometry, BD, Triangle, NC, USA) for PE-Cy7 conjugated anti-mouse CD34 monoclonal antibody.

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acetylated low-density lipoprotein (DiI-ac-LDL; Maokang, China) in EGM-2 medium for 4 h at 37 °C. Cells were fixed in 4% paraformaldehyde.

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Dil-Ac-LDL/ FITC-UEA-1 staining

The adherent cells were incubated in a medium containing Dil-Ac-LDL (2.4 μg/ml; MKBio, Shanghai, China) at 37 °C for 1 h, washed twice, and fixed in 4% paraformaldehyde for 10 min. The cells were then incubated in FITC-UEA-1 lectin (10 μg/ml; MKBio, Shanghai, China) for 1 h. 4', 6-diamidino-2-phenylindole (DAPI, Beyotime Biotechnology, Shanghai, China) was used to observe the cell nuclei. Images were acquired using a confocal microscope (Nikon, Tokyo, Japan). Double-positive staining for DiI-Ac-LDL and FITC-UEA-1 lectin was considered an EPC.

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FITC-UEA-1 was purchased from Maokang Biotechnology Co., Ltd (Shanghai, China).

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Later, 2% paraformaldehyde was used to fix the cells and FITC labeled Ulex europaeus agglutinin 1 (FITC-UEA-1) (Shanghai Maokang Biotechnology Co., Ltd, China) to incubate the