Dil-Ac-LDL – Cell Labeling Reagent (100ug)

Biomedical Technologies Inc

NOTE : THIS PRODUCT HAS ERRATIC AVAILABILITY PLEASE CALL FIRST BEFORE ORDERING.

DiI-Ac-LDL Reagent

  • Labels Endothelial Cells & Macrophages
  • DiI Labeled Cells can be Trypsinized
  • Does Not Label Matrix Associated Proteins
  • DiI Labeled Cells Remain Viable
  • Directly Applicable to Cell Sorting & Fluorescence Microscopy

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.

Procedural Outline

1. Dilute DiI-Ac-LDL to 10ug/ml in complete 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.

References

1. Goldstein, J.L., Y.K., Ho. S.K. Basu, and M.S. Brown. 1979. Binding site on macrophages that mediates uptake degradation of actylated low density lipoprotein, producing massive cholesterol deposition. Proc. Nat. Acad Sci. USA 76: 335-337.

2. Folgelman, A.M., I. Schechter, J. Seager, M. Hokum, J.S. Child and P.A. Edwards. 1980. Malondialdehyde alteration of low density lipoproteins leads to cholesterylester accumulation in human monocyte-macrophages. Proc. Nat. Acad. Sci 77: 2214-2218.

3. Stein, O. and Y. Stein 1980. Bovine aortic endothelial cells display macrophage-like properties towards acetylated [I125]-labeled low density lipoprotein. Biochem. Biophys. Acta 620: 631-635.

4. Pitas, R.E., T.L. Innerarity, J.N. Weinstein, and R. W. Mahley. 1981. Acetoacetylated lipoproteins used to distinguish fibroblasts from macrophages in vitro by fluoresence microscopy. Arteriosclerosis. 1:  177-185.

5. Voyta, J.C., D.P. Via,  C.E. Butterfield and B.R. Zetter. 1984.  Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J. Cell Biology. 99:  2034-2040.

6. Giulian, D. and D. G. Young. 1986. Brain peptides and glial growth. II. Identification of cells that secrete glia-promoting factors. J. Cell Biology. 102:  812-820.

7. Netland, P.A., B.R. Zetter, D.P. Via and J. C. Voyta. 1985. Insitu labeling of vascular endothelium with fluorescent acetylated low density lipoprotein. Histochemical Journal. 17: 1309-1320.

8. Pitas, R.E., J. Boyles, R. W. Mahley and D.M. Bissell. 1985. Uptake of chemically modified low density lipoproteins in-vivo is mediated by specific endothelial cells. J. Cell Biology. 100: 103-117.

9. Giulian, D., and T. J. Baker 1986. Characterization of ameboid microglia isolated from developing mamalian brain. J. Neuroscience.  6:  2163-2178.

10. Bjorling, D.E., R. Saban, M.W. Tengowski, S.M. Gruel and V.K. Rao. 1992. Removal of Venous Endothelium with Air. J. Pharm. & Tox. Methods. 28: 149-157.

11. Lysco, P.G., J. Weinstock, Chris. T. Webb, M.E. Brawner and N.A. Elshourbagy. 1999. Identification of a Small-Molecule, Nonpeptide Macrophage Scavenger Receptor Antagonist. J. of Pharm. and Experimental Therapeutics. 289 No.3: 1277-1285.

12. Murphy, H.S., R.L.Warner, N. Bakopoulos, M.K. Dame, J. Varani and P.A. Ward. 1999. Endothelial Cell Determinants of Susceptibility to Neutrophil-Mediated Killing. Shock 12:111-117.

13. Nugent, M. A. et al. 2000. Perlecan is required to inhibit thrombosis after deep vascular injury and contributes to endothelial cell-mediated inhibition of intimal hyperplasia. PNAS. 97 No.12: 6722-6727.
Catalog Information

DiI-Ac-LDL
Catalog No:  BT-902
Quantity:  100ug
(Including labeling protocol)

Refrigerate at 4ºC.  Do Not Freeze.

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS OR AS AN IN-VITRO DIAGNOSTIC.

Dil-Ac-LDL – Cell Labeling Reagent (100ug)
Shipping Standard

Buy now

£535.00 / €749.00 BT-902 (Dil-Ac-LDL) (100ug)

All prices shown are exclusive of VAT