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BTI Lipoproteins/Proteins
BTI Lipoproteins
are isolated from fresh blood bank produced human plasma. Each product
is prepared via ultracentrifugation and is analyzed by agarose gel electrophoresis.
Most of our lipoproteins are evaluated in conjunction with our [I125]
or DiI labeled lipoproteins in cell receptor/lipoprotein metabolism studies.
Our fluorescently labeled DiI-Ac-LDL and DiI-LDL conveniently label (in
one step) cultured cells for identification, isolation or quantification
of receptor site activity. Most of our lipoproteins are membrane filtered
and aseptically filled. Some of our Lipoproteins are Endotoxin tested.
Quantities listed are milligrams or micrograms of Protein. BTI prepares Apolipoproteins
and a few other proteins, Tamm-Horsfall Glycoprotein and antibodies.
ABBREVIATIONS:
Human
VLDL Apolipoproteins:
Apo A-I and Apo A-II Human HDL BTI Human High Density Lipoprotein, HDL, is isolated from blood bank produced human plasma. It is purified via ultracentrifugation (1.063-1.21g/cc) to homogeneity determined on agarose gel electrophoresis. BTI HDL is aseptically packaged and should be stored at 4oC. Human HDL [I125]
Human HDL anti-LDL IgG Immunohistochemical Grade BTI anti-Human Low Density Lipoprotein (LDL) is prepared in rabbits using BTI purified LDL immunogen. The resulting serum is affinity purified on an LDL sepharose column. Both Human LDL and VLDL react with the antibody but not human HDL as determined via immunodiffusion and Elisa methods. The antibody is suitable for the immunohistochemical localization of LDL, Ac-LDL and Ox-LDL in cells or tissues. Each lot is analyzed for the localization of LDL or Ac-LDL receptors by ligand blotting procedures. anti-Human LDL Ox-LDL, DiI-OxLDL and [I125] Ox-LDL BTI Oxidized Human LDL is made via copper sulfate oxidation. Each lot is analyzed for the degree of oxidation (via TBARS assay) and migration versus the native LDL on agarose gel electrophoresis. The product is membrane filtered, aseptically packaged and Endotoxin tested. Our Ox-LDL is evaluated ultimately for receptor binding to peritoneal macrophages in conjunction with our [I125] Ox-LDL or DiI-Ox-LDL. These Ox-LDL products have a short shelf-life, plan your experiments accordingly. Human Ox-LDL Human Ox-LDL (Hi TBAR Ox-LDL)
DiI-Ox-LDL Lipoprotein Deficient Serum BTI Lipoprotein Deficient Serum, LPDS, is prepared by ultracentrifugation, all of the lipoproteins are removed and it is extensively dialyzed. The resultant LPDS is membrane filtered and packaged aseptically. Each lot is evaluated in Human Skin Fibroblast or other cells in culture with our LDL and [I125] LDL (or DiI-LDL and DiO-LDL) to determine the degree of up regulation of LDL receptors. Typical lots give a 3 to 4 fold increase in [I125] LDL binding. We do not dilute the LPDS, typical protein concentration for the bovine LPDS is 75 -120 mg/ml. and for the human LPDS is ~50mg/ml. Bovine LPDS Catalog No: BT-907
* Quantity: On Request Reference
DiI-HDL DiO-LDL, Cell Marker
Human Lipoprotein [a], Lp
[a] anti-Human Apo [a] (LDL
Adsorbed) DiI-Ac-LDL & LDL Reagents
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. 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 Refrigerate at 4ºC. Do Not Freeze. DiO-Ac-LDL Refrigerate at 4ºC. Do Not Freeze. FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS OR AS AN IN-VITRO DIAGNOSTIC. Human LDL & Ac-LDL BTI Human Low Density Lipoprotein, LDL, is isolated from blood bank produced human plasma. It is purified via ultracentrifugation to homogeneity determined by agarose gel electrophoresis. Each lot of BTI LDL is evaluated for receptor binding to human skin fibroblasts in conjunction with our [I125] LDL or DiI-LDL. Acetylated Human Low Density Lipoprotein, Ac-LDL, is also available. The BTI Ac-LDL is purified to homogeneity as demonstrated on agarose gel electrophoresis. Each lot of BTI Ac-LDL is evaluated for receptor binding to murine peritoneal macrophages in conjunction with our [I125] Ac-LDL or DiI-Ac-LDL. LDL, Human Ac-LDL, Human [I125] Human Ac-LDL
Human Ac-LDL (99% Pure, AGE) is radioiodinated by the Iodine Monochloride Method. The resulting [I125] Ac-LDL is purified via column chromatography. Each lot is evaulated for binding to murine peritoneal macrophages. [I125]
Human Ac-LDL [I125] Human LDL Human LDL (99% Pure, AGE) is radioiodinated by the Iodine Monochloride Method. The resulting [I125] LDL is purified via column chromatography. Each lot of BTI [I125] LDL is evaluated for binding to human skin fibroblasts.
DiI-LDL, Cell Marker
DiI-LDL, Cell Marker
DiO-LDL(FITC-like) References 1. Pitas R.E.,
et al. Arteriosclerosis 1: 177 (1983). Other Proteins Calmodulin BTI Calmodulin is purified to >99.5% as demonstrated by silver stained, two dimensional polyacrylamide gel electrophoresis. The biological activity is 48,000 units/mg (one unit of Calmodulin gives a 50% activation of 0.14 units of bovine retina adenylate cyclase where one unit of adenylate cyclase is equivalent to the production of one nanomole of Cyclic AMP per minute per milligram of protein at 37ºC). This product is suitable for radioiodination and as a standard in biological and immunohistochemical experiments. Catalog No: BT-372 References
Tamm-Horsfall Glycoprotein Tamm-Horsfall Glycoprotein, Uromodulin, is the most abundant protein present in normal human urine. Recent studies have implicated THG as a unique regulatory glycoprotein limiting the circulating activity of a number of potent cytokines. BTI THG is purified from normal human urine via precipitations with sodium chloride and then by Sepharose gel chromatography. The protein shows a single homogeneous band on 5-18% PAGE run under reducing and non-reducing conditions. BTI THG is packaged in aqueous solution with a stabilizer. Catalog No: BT-381 anti-Tamm-Horsfall Glycoprotein BTI anti-Human Tamm-Horsfall Glycoprotein is prepared in rabbits using chromatographically pure THG. No significant cross-reactivity with urinary or other related proteins was detected. This antibody is suitable for Elisa assays (1:150,000 against solid phase antigen) and localization studies on cells and tissues (1:100 to 1:250). Catalog No: BT-590 References 1. I. Tamm
and F.L. Horsfall, Proc. Soc. Exp. Biol. Med. 74: 108-114 (1950).
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