Application

E06 mAb-TopFluor^® has been used: in the detection of oxidized phospholipids by immunocytochemical method in human primary skin fibroblastsas a component of blocking solution in competitive binding assay of oxidized phosphatidylcholinefor the detection of oxidized phospholipids (OxPL) in the extracellular vesicle by flow cytometry

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.

Biochem/physiol Actions

E06 blocks the transient receptor potential cation channel subfamily V member 1 (TRPV1) channels.

General description

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. 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.

E06, an IgM mouse autoantibody is also called T15 derived from apolipoprotein E‐deficient mice.

Legal Information

TopFluor is a registered trademark of Avanti Polar Lipids, Inc.

Packaging

0.5 mL Polypropylene Tube with Screw Cap (330003S-25ug)

Preparation Note

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.