DiI-LDL
$2,600.00
| Name: | DiI-LDL | ||
| Cat. #: | 10459 | ||
| Size: | 1.0 mg | ||
| Purity: | 98% (Co-migrates with reference on agarose gel electrophoresis) | ||
| Concentration: | Minimum 1.6 mg/ml protein | ||
| Description: | Human DiI-Labeled Low Density Lipoprotein | ||
| Background: | Purified LDL is labeled with the fluorescent probe, DiI, and reisolated by ultracentrifugation (1.019-1.063). The resultant product is exhaustively dialyzed against phosphate buffered saline, (pH 7.4), sterilized by membrane filtration and then aseptically packaged in a solution containing phosphate-buffered saline at pH 7.4 and 0.2 mM EDTA. Each lot is evaluated on a murine macrophage cell line for fluorescence uptake. | ||
| Storage & Stability: | The labelled LDL is stable for 6 weeks after receipt when handled aseptically and stored at 2-8°C (Don’t Freeze). Note: After prolonged storage, some precipitate may be observed. This is normal for the product. Spin in centrifugation at 5000×g for 10 minutes before using. | ||
| Packaging: | The labelled LDL requires one week lead time. Please plan your experiments in advance and use the fresh material. | ||
Native-LDL(n-LDL), Oxidized-LDL (ox-LDL) and -LDL(Ac-LDL) were loaded on agarose gel and electrophoresed for 60 mins. The lipoproteins were stained with Sudan Black (A and B). Oil red O staining was used to determine the formation of foam cell. RAW264.7 were incubated with 80 μg/mL ox-LDL for 24 hrs.
Typical Lipoprotein Labeling Protocol
- Dilute DiI-LDL to 10-40 ug/ml in growth media.
- Add to cells and incubate for 2-6 hours at 37ºC.
- Remove media containing DiI-LDL from your culture.
- Wash 3 times with probe-free media.
A. Fluorescence Microscopy: Visualize using standard rhodamine excitation: emission filters (or suggested wavelengths excitation:emission at 554nm:571nm or near). 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.
A. 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: 554nm
Emission: 571nm
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