北京索莱宝科技有限公司  张净花  010-56371206   13426459985


DiI-Ac-LDL DiI标记乙酰化低密度脂蛋白
品牌：Solarbio | 货号：H7970 | CAS：


 


    
储存条件 : 2-8℃避光保存，请勿冷 冻
    
纯度 : 98%

 

 

    

        

            
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H7970-500ug
            
Solarbio
            
500ug
            
2500.00元
        
    

产品介绍
DiI-Ac-LDL，即1,1’-双十八酯基-3,3,3,,3,四甲基-吲哚碳菁高**盐标记的乙酰化低密度脂蛋白。它能用于鉴定和分离混合细胞群中的血管内皮细胞和巨噬细胞。当细胞被DiI-Ac-LDL标记后，脂蛋白就会被溶酶体酶降解并且DiI（荧光探针）会积累在细胞内膜中。标记了DiI-Ac-LDL的细胞生存活性不受影响。血管内皮细胞纯化培养物可以基于增加的代谢DiI-Ac-LDL利用荧光激活分类从复杂的初级培养物中进行分离。污染细胞（成纤维细胞、平滑肌细胞、周细胞、上皮细胞）将不会被标记。巨噬细胞能够从混合的细胞群中（包括血管内壁细胞）区分出来是因为它的标记更亮一些。
      标记有DiI-Ac-LDL的内皮细胞较其它抗原标记的内皮细胞有较多的优势。一旦细胞被标记， 荧光探针(DiI)将不会被胰蛋白酶移除掉。低密度和汇合培养的血管内皮细胞都将被高效的标记。其它类型的细胞标记水平均达不到血管内皮细胞标记（除了巨噬细胞）。索莱宝公司DiI-Ac-LDL均通过牛主动脉内皮细胞和小鼠巨噬细胞进行标记测验以确保结果的一致性。
实验步骤：
      1. 在生长培养基中将DiI-Ac-LDL稀释至20-50ug/ml；
      2. 将其加入到细胞中37 ºC温育4小时；
      3. 去掉培养基；
      4. 用无探针的缓冲液冲洗；
      5. 通过荧光显微镜观察并/或者将细胞通过胰蛋白酶（或者EDTA）进行分类。
      6. 荧光显微镜：用标准罗丹明激发进行观察 （或者建议用波长激发：549nm、发射：565nm）。如果需要，用3%甲醛在PBS中固定。切勿使用甲醇或丙酮进行固定，因为DiI溶于有机溶剂。（仅供科研使用）
注意事项：
长时间的储存后可能会产生沉淀，这种现象属于正常情况。溶液放入离心管中离心2分钟将沉淀除去即可。DiI-Ac-LDL不稳定，实验时*好现用现配，采用新鲜配置工作液。
DiI-LABELED ACETYLATED LOW DENSITY LIPOPROTEIN, HUMAN
Catalog No: H7970
Lot No: 2015-10-12
Quantity: 500ug(micrograms)Protein/Vial
Concentration: 1.6mg/ml (Protein)
Storage:
This product is stable for 6 weeks when handled aseptically and stored at 2-8°C. protect from light and never freeze.
Introduction
DiI-Ac-LDL, Acetylated Low Density Lipoprotein, labeled with 1,1-dioctadecyl – 3,3,3,3-tetramethyl-indocarbocyanine perchlorate, labels both vascular endothelial cells and macrophages. It can be used to identify and/or isolate these cells from mixed cell populations. When cells are labeled with DiI-Ac-LDL, the lipoprotein is degraded by lysosomal enzymes and the DiI (fluorescent probe) accumulates in the intracellular
membranes. Labeling cells with DiI-Ac-LDL has no effect on cell viability. Pure cultures of vascular endothelial cells can be isolated from complex primary cultures using fluorescent activated cell sorting based on their increased metabolism of the DiI-Ac-LDL. Contaminating cell types (fibroblasts, smooth muscle,pericytes, epithelial cells) are not labeled. Macrophages can be differentiated from mixed cell populations (including endothelial cells) because they are more brightly labeled.
Labeling endothelial cells with DiI-Ac-LDL has many advantages over labeling other endothelial cell associated antigens. The labeling procedure is one step, and once the cells are labeled, the fluorescent probe (DiI) is not removed by Trypsin. Both low density and confluent cultures of vascular endothelial cells are effectively labeled. No other cell type (other than macrophages) is labeled to the same level as vascular endothelial cells. Each lot of DiI-Ac-LDL is evaluated for the specific labeling of bovine aortic endothelial
cells and murine macrophages to assure consistent results. A complete labeling protocol is included with each shipment. We also offer an "FITC-like" label DiO-Ac-LDL, which is useful for fixed wavelength FACS Cell sorters.
Special Note:
After prolonged storage, some precipitate may be observed. This is normal for this product. Clarify out the
aggregates by spinning in a microfuge for 2 minutes.
Preparations of DiI-Ac-LDL are fairly unstable; plan your experiments in advance and use fresh material.
Procedural Outline
1. Dilute DiI-Ac-LDL to 20-50ug/ml in growth media.
2. Add to cells and incubate for 4 hours at 37ºC.
3. Remove media.
4. Wash with probe-free media.
5. Visualize via Fluorescence Microscopy and/or trypsinize (or EDTA) for cell sorting.
6. Fluorescence Microscopy:
Visualize using standard rhodamine excitation: emission filters (or suggested wavelengths
excitation:emission at 549nm:565nm). If fixation is desired use 3% formaldehyde in PBS. (Never use
methanol or acetone fixation - DiI is soluble in organic solvents).
Note: 
A.positive culture must be stained for comparison purposes.
B. Cell Sorting:Label as in steps 1-5. Trypsinize or treat cultures with EDTA to produce a single cell suspension.Use labeled pure cultures of positive and negative cell types to set gates on the cell sorter.
Suggested Wavelengths for Cell Sorting: Excitation: 514/549nm；Emission: 565nm
 
北京索莱宝科技有限公司  张净花  010-56371206   13426459985
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因为此类产品保质期较短，所以都是现做的，订购后1-2个工作日可以发货