文献支持DiI标记氧化型低密度脂蛋白 DiI-Ox- LDL

保存条件 ：RT/2-8℃/-20℃

保质期 ：3年

英文名 ：DiI-LLabeled oxidized low density lipoprotein, human

库存 ：999

供应商 ：北京康瑞纳生物科技有限公司

CAS号 ：152-1055-9202

规格 ：0.5mg

DiI标记氧化型低密度脂蛋白 DiI-Ox- LDL DiI标记氧化型低密度脂蛋白 DiI-Ox- LDL
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产品名称：DiI-Ox- LDL DiI标记氧化型低密度脂蛋白
产品编号：R1050
中文别称 DiI标记氧化型低密度脂蛋白
英文别称 DiI-LLabeled oxidized low density lipoprotein, human
储存条件 2-8℃避光保存，请勿冷冻。
有效期 6周
来源：人
浓度：1.0mg/ml
纯度 98%
规格 0.5mg
DiI labeled oxidized low-density lipoprotein DiI-Ox LDL
Product name: DiI-Ox LDL DiI labeled oxidized low-density lipoprotein
Product number: R1050
Chinese alias DiI labeled oxidized low-density lipoprotein
English alias DiI LLAbeled oxidized low density lipoprotein, human
Storage conditions: 2-8 ℃, avoid light and do not freeze
Validity period of 6 weeks
Source: People
Concentration: 1.0mg/ml
Purity 98%
Specification 0.5mg
Oxidized low-density lipoprotein, labeled with 1,1 '- didecyl-3,3,3' -, 3 '- tetramet.hylindocyanine perchlorate, is a highly fluorescent lipophilic dye that can diffuse to the hydrophobic part of LDL complexes without affecting the LDL specific binding of apolipoprotein. DiI-Ox-LDL is used to label endothelial cells and macrophages. It can be used to identify and/or isolate these cells from mixed cell populations, and to investigate the uptake and binding of modified LDL by different cell types. DiI is excited at 488nm or 514nm and emission is detected at 565nm.
Dil Ox low-density lipoprotein Dil labeled oxidized low-density lipoprotein
Packaging: 0.5mg protein/small bottle
Concentration:>1 mg/mL protein (based on actual label concentration)
Storage:
When sterile and stored at 2-8 ° C, this product can remain stable for 6 weeks upon receipt (do not freeze). After long-term storage, some precipitates may be ob.served. This is normal for this product. Clarify the aggregates by rotating them in a centrifuge tube for 2 minutes.
Introduction:
Dil Ox LDL, oxidized low-density lipoprotein, labeled endothelial cells and macrophages with 1,1 '- octadecyl-3,3,3' -, 3 '- tetramet.hyl-perchlorite indocyanine.
It can be used to identify and/or isolate these cells from mixed cell populations, and to investigate the uptake of modified LDL by different cell types.
When cells are labeled with Dil-Ox-LDL, lipoproteins are accumulated in the cell membrane by lysosomal degrading enzymes and Dil (fluorescent probe). Labeling cells with Dil Ox LDL has no effect on cell viability.
The pure culture of vascular endothelial cells can be obtained from complex primary cultures sorted using fluorescent activated cells based on Dil-Ox-LDL metabolism.
The types of contaminated cells (fibroblasts, smooth muscle cells, pericytes, epithelial cells) are not labeled.
Macrophages can differentiate from mixed cell populations (including endothelial cells) because they are labeled brighter. Compared to labeling other endothelial cell related antigens, labeling endothelial cells with Dil-Ox-LDL has many advantages. The labeling program is a step, once a cell is labeled
The fluorescent probe (Dil) is not removed by trypsin. The low density and fusion cultured endothelial cells of blood vessels are effectively labeled. No other cell types (excluding macrophages) were labeled to the same level as vascular endothelial cells.
Evaluate the specific labeling of each batch of Dil-Ox-LDL on bovine aortic endothelial cells and mouse macrophages to ensure consistent results.
1. Dilute Dil OxLDL sterile to 20-40 in the culture medium μ G/mL.
2. Add to live cells and incubate at 37 ℃ for 3-6 hours.
3. Remove the medium containing Dil OxLDL from the culture medium.
4. Wash cells several times with probe free culture medium.
5. A. Fluorescence microscope:
Use standard Rhodamine excitation: emission filter (or recommended wavelength excitation: emission) for visualization at 549nm: 565nm). If fixation is required, use 3% formaldehyde in PBS. (Do not use methanol or ace.tone to fix - Sil is soluble in organic solvents).
Note: Positive cultures must be stained for comparison.
B. Cell sorting
Mark according to steps 1-5. Phosphorylate or treat the culture with EDTA to produce a single-cell suspension. Set gates on the cell classifier using labeled pure cultures of positive and negative cell types.
Suggested wavelength for cell sorting: excitation: 514/549nm; Emission: 565nm
Fixation and installation of Dil labeled cells
Wash three times in PBS.
2. Fix at room temperature in 3% formaldehyde/PBS for 20 minutes.
3. Rinse with distilled water at room temperature for 5 seconds.
4. Discharge the liquid onto a chemical cloth.
5. Invert the lid and drop 90% glycerol and 10% PBS onto a microscope slide.
6. Seal with Kroenigs wax. Do not use nail polish. The storage temperature is -20 ℃.
Special note: Low density lipoprotein products tend to naturally aggregate. The aggregates of this product may interfere with its use. To understand these aggregates, simply rotate them in a micro centrifuge for 2 minutes.
Representative confocal images of DiI ox LDL uptake in rats - phages from ApoE knockout mice.

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