DiI-LDL DiI标记低密度脂蛋白

Dil-Low Density Lipoprotein DiI标记低密度脂蛋白

Low Density Lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate

Cat No:C02650

Concentration:>1mg/mL protein(Based on actual label concentration)Buffer containing phosphate- buffered saline at pH 7.4 and 0.2 mM EDTA.

Specifications:0.5mg/vial;0.22 micron membrane filtered,aseptically filled.Cell Culture Tested. Absorbance Ratio:DiI/Protein=555nm/275nm=1.40

Storage

This product is stable for 6 weeks after receipt when handled aseptically and stored at 2-8℃(Do not free). After prolonged storage,some precipitate may be observed.This is normal for this product.Clarify out the aggregates by spinning in centrifuge tube for 2 minutes.LDL products have a natural tendency to aggregate.Aggregates of this product can interfere with its use.

Product Preparation:

Purified LDL is labeled with the fluorescent probe,Dil,and reisolated by ultracentrifugation(1.019-1.063).

Each lot is evaluated on a murine macrophage cell line for fluorescence uptake.

Typical Lipoprotein Labeling Protocol

1. Aseptically dilute the Dil-LDL to 10-40 μg/mL in growth media.
2. Add to live cells and incubate for 4-6 hours at 37℃.
3. Remove media.
4. Wash cells several times with probe-free media.
5. 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.(Do not use methanol or acetone fixation-Dil is soluble in organic solvents).

Note:A positive culture must be stained for comparison purposes.

1. 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.

Fixation and Mounting of DiI Labeled Cells

1. Wash 3 times in PBS.
2. Fix in 3%formaldehyde/PBS for 20 minutes at room temperature.
3. Rinse 5 seconds in distilled water at room temperature.
4. Drain liquid onto chem-wipe.
5. Invert cover,slip on a drop of 90%Glycerol and 10%PBS onto a microscope slide.
6. Seal with Kroenigs wax,also known as cover glass cement Do not use nail polish.Store at -20℃.