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E06 mAb-TopFluor™ | 330003

E06 Mouse Monoclonal Antibody (IgM), TopFluor® labeled
Anti-(Oxidized Phospholipid)

330003S

Saline

25 μg
$245.00

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The E06-TopFluor® is primarily use for direct ELISA, immune-fluorescence microscopy and flow cytometry.
 
The E06 antibody is a murine monoclonal IgM natural antibody that binds to oxidized phospholipid and reacts with oxidized LDL, oxidized HDL and protein covalently modified by oxidized phospholipid. This antibody has been used to probe atherosclerotic plaque in mice, zebrafish, rabbits, monkeys and humans. It also binds to apoptotic cells, but not viable cells, and can be used in a variety of applications, including ELISA, immunohistochemistry and Western blot analysis, as well as for in vivo imaging with MRI techniques.

Avanti now provides high purity E06 from animal-free culture of hybridoma cells. Each batch is verified for activity and provided with detailed protocols for the above applications.

The E06 antibody was developed by Dr. Joseph Witztum at the University of California, San Diego using apolipoprotein E knock-out mice fed a high-fat diet for seven months.
Purity
>95%
Stability
6 Months
Storage
We recommend long-term storage of E06 at -80°C. After it has been thawed for initial use, we recommend storing at 4°C. We do not recommend freeze-thawing or aliquoting.
Synonyms
T15 antibody
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Notes
Palinski W, Hörkkö S, Miller E, Steinbrecher UP, Powell HC, Curtiss LK, Witztum JL. Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice. Demonstration of epitopes of oxidized low density lipoprotein in human plasma. J Clin Invest. 1996 98: 800-14.


Tsimikas S, Aikawa M, Miller FJ Jr, Miller ER, Torzewski M, Lentz SR, Bergmark C, Heistad DD, Libby P, Witztum JL. Increased plasma oxidized phospholipid:apolipoprotein B-100 ratio with concomitant depletion of oxidized phospholipids from atherosclerotic lesions after dietary lipid-lowering: a potential biomarker of early atherosclerosis regression. Arterioscler Thromb Vasc Biol. 2007 27: 175-81.


Hörkkö S, Bird DA, Miller E, Itabe H, Leitinger N, Subbanagounder G, Berliner JA, Friedman P, Dennis EA, Curtiss LK, Palinski W, Witztum JL. Monoclonal autoantibodies specific for oxidized phospholipids or oxidized phospholipid-protein adducts inhibit macrophage uptake of oxidized low-density lipoproteins. J Clin Invest. 1999 103: 117-28.



Friedman P, Hörkkö S, Steinberg D, Witztum JL, Dennis EA. Correlation of antiphospholipid antibody recognition with the structure of synthetic oxidized phospholipids. Importance of Schiff base formation and aldol condensation. J Biol Chem. 2002 Mar 1;277(9):7010-20.


Chang MK, Bergmark C, Laurila A, Hörkkö S, Han KH, Friedman P, Dennis EA, Witztum JL. Monoclonal antibodies against oxidized low-density lipoprotein bind to apoptotic cells and inhibit their phagocytosis by elicited macrophages: evidence that oxidation-specific epitopes mediate macrophage recognition. Proc Natl Acad Sci U S A. 1999 96: 6353-8.

For binding studies with the E06-TopFluor™ antibody, 96-well Microfluor 1 black U bottom plates (Thermo 7005) were coated with 50µL of antigen (5µg/mL) in PBS containing 0.27 mM EDTA and overnight at 4°C. The wells were washed three times with PBS containing 0.27 mM EDTA. After washing, the plates were blocked with 3% BSA in PBS for 45 min at room temperature. 100µL aliquots of a serial dilution of the E06-TopFluor™ antibody (0.78-400ng/well) in PBS containing EDTA and 1% BSA were added to each well and incubated 1hr at room temperature. The amount of IgM bound to each well was quantified with after 1hr incubation at room temperature with excitation (Ex) at 485 nm and emission (Em) recorded at 520 nm.