HNE (4-Hydroxy-2-nonenal), Western Blot Control (BSA)

HNE is a major product of endogenous lipid peroxidation. The w-6-family (linoleic and arachidonic acids) of polyunsaturated fatty acids may produce HNE as a result of free radical attack. HNE is a highly reactive compound and it can react with several functional groups on biological material, particularly sulfydryl groups to form thioester adduct and then hemiacetals. HNE may also react with histidine and lysine residues of proteins to form stable Michael addition-type of adducts. HNE-modified proteins may display an altered biological functions. Antibodies to HNE will help to visualize the HNE-adducts.\n\nApplications: \nSuitable for use in Western Blot as positive control. Not suitable for use in ELISA. Other applications have not been tested.\n\nRecommended Dilution:\nWestern Blot: 10 ul/lane. Loading volume may be adjusted depending upon the intensity of the band under actual experimental condition. BSA-HNE conjugate is ~65-70kD. Other aggregated bands may be visible as well. BSA-HNE conjugate should only be used as a positive control to optimize the blotting conditions. It is not a representative of the protein bands that might be labeled with the anti-HNE in a given samples. Typically, complex proteins samples, such as liver or brain, will show many bands that are modified with HNE. There is no specific pattern that must be obtained with the antibodies.\n\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

H6276-05A

Size

100ul

Applications

WB

Form

Supplied in 100ul of SDS-PAGE sample buffer (reduced).

Purity

Purified

References

1. Yoritaka, A., et al., Proc. Natl. Acad. Sci. 93: 2696-2701 (1996). 2. Uchida, K., Arch. Biochem. Biophys. 324: 241-248 (1995). 3. Quinn, M.T., et al., J. Leukocyte Biol. 57: 415-421 (1995). 4. Okamoto, K., Int. J. Cancer 58: 825-829 (1994). 5. Uchida, K., et al., Proc. Natl. Acad. Sci. 90: 8742-8746 (1993).