Advanced BioDesign can support you to develop and produce rabbit polyclonal antibody offering you a taylor-made & ready-to-use tool.

To discover our services, we offer 10% discount on Polyclonal Package in February 2012 until March 2nd.

For more information please contact: contact@a-biodesign.com

Nowadays, oxidative stress may involved in cell perturbation leading to various diseases like atherosclerosis, heart failure, myocardial infarction, Alzheimer’s disease, schizophrenia…

The six consecutive histidine residues take place on the N or C terminus of protein. This amino acid will form an electronegative cloud that can be bind by positively charged metal ions such as nickel. This linkage can be broken using high concentration of imidazole result the release of the tagged protein.

This product may be used for monitor His-tagged protein such as expression screening, immunoassays, competition assays, biochemical assays or protein purification.

The limit of detection of this product is 10pg of His-tagged protein per well.

Highlights

• Detection limit: 10pg of histidine tagged protein.
• Detergents do not interact with the binding capacity of the plate.
• Pre-blocked plate for reducement of non-specific binding
• Use optimizes Wash Buffer for lower background.
• Can be used for little purification of histidine-tagged protein.
• This plate can be chelated with other metals ions. Please contact our technical services for any request.

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Highlights

• Ready to use HRP Substrate
• Sensitivity (ng)

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Using this new substrate is also a reliable low cost solution for enhanced chemiluminescence (ECL) that will replace more expensive products without losing sensibility.

ABD uses in this substrate a new luminol enhancer for boosting signals without increasing the background.

Highlights

• Low cost ECL Substrate: cost less than similar chemiluminescent substrates.
• New Luminol Enhancer: switching to ABD ECL substrate for Western blot detections yielding strong signals and low background.
• Saving antibody: requires less primary and/or secondary antibodies per blot.

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Advanced BioDesign Team

Oxidative damage of lipids caused by reactive oxygen species (ROS) play an important role in some diseases, lesion of cell functions and aging. Aldehydes such as malondi-aldehyde (MDA) and 4-hydroxy-2-nonenal (4-HNE) have been reported as one of the advanced lipid peroxidation products. But recently in the earlier stage of lipid peroxidation, 13-hydroperoxyoctadecanoic acid (13-HpODE) is found to be covalently bound to proteins. Hexanoyl-Lysine adduct (HEL) is a novel lipid hydroperoxide-modified lysine residues. HEL is formed by oxidative modification by oxidized omega-6 fatty acids such as linoleic acid or arachidonic acid. This monoclonal antibody (clone 5H4)is spesific for proteins and peptides containing HEL. Suitable for HEL detection by immunohistochemistry and western blotting.

Product Information

• 02-0702: Anti Hexanoyl-Lys(HEL) monoclonal antibody (clone 5F12) (20µg)

Product Specification

• Clone: 5F12
• Immunogen: N-ε-Hexanoyl modified keyhole limpet hemocyanin (KLH)
• Specificity: Specific to hexanoyl adduct. Cross reactivity has been checked for following oxidized lipids: MDA, glyoxal, methylglyoxal, 1-hexanal, 2-hexenal, 1-nonnanal, 2-nonenal & HNE
• Subclass: Mouse IgG1(κ)
• Form: Lyophilized powder, containing 100mM PBS, 5% sucrose, 1% BSA
• Applications: Immunohistochemistry (recommended concentration 2µg/mL)
• Storage: Store at less than  -20°C

Detection of HEL at Human atherosclerotic lesions.

References

• Yoji Kato, Yoko Mori, Yuko Makino, Yasujiro Morimitsu, Sadayuki Hiroi, Toshitsugu Ishikawa, Toshihiko Osawa, Formation of N epsilon-(hexanonyl) lysine in protein exposed to lipid hydroperoxide. J Biol Chem 274(29), p20406-20414 (1999)
• Kato Y, Miyake Y, Yamamoto K, Shimomura Y, Ochi H, Mori Y, Osawa T, Preparation of a monoclonal antibody to N(epsilon)-(Hexanonyl)lysine: application to the evaluation of protective effects of flavonoid supplementation against exercise-induced oxidative stress in rat skeletal muscle. Biochem Biophys Res Commun 274(2),p389-393(2000)
• Fukuchi Y, Miura Y, Nabeno Y, Kato Y, Osawa T, Naito M, Immunohistochemical detection of oxidative stress biomarkers, dityrosine and N(epsilon)-(hexanoyl)lysine, and C-reactive protein in rabbit atherosclerotic lesions. J Atheroscler Thromb 15(4)p185-192(2008)
• Maeda R, Noiri E, Isobe H, Homma T, Tanaka T, Negishi K, Doi K, Fujita T, Nakamura E, A water-soluble fullerene vesicle alleviates angiotensin II-induced oxidative stress in human umbilical venous endothelial cells. Hypertens Res 31(1)p141-151(2008)
• Sango K, Yanagisawa H, Kato K, Kato N, Hirooka H, Watabe K: Differential Effects of High Glucose and Methylglyoxal on Viability and Polyol Metabolism in Immortalized Adult Mouse Schwann Cells. Open Diabetes J. 1, p1-11(2008)

Oxidative damage of lipids caused by reactive oxygen species (ROS) play an important role in some diseases, lesion of cell functions and aging. Aldehydes such as malondi-aldehyde (MDA) and 4-hydroxy-2-nonenal (4-HNE) have been reported as one of the advanced lipid peroxidation products. But recently in the earlier stage of lipid peroxidation, 13-hydroperoxyoctadecanoic acid (13-HPODE) is found to be covalently bound to proteins1). Hexanoyl-Lysine adduct (HEL) is a novel lipid hydroperoxide-modified lysine residues. HEL is formed by oxidative modification by oxidized omega-6 fatty acids such as linoleic acid or arachidonic acid. HEL may be a useful biomarker for initial stage of lipid peroxidation.Monoclonal antibodies and ELISA kit have been developped, and HEL can be detected in oxidatively modified LDL, in human atherosclerotic lesions, human urine and serum. It is also reported that HEL is formed in rat muscle during exercise, and the formation is prohibited by antioxidants such as flavonoids.

Hexanoyllsyine formation process

Product Information

• 02-0701: Hexanoyl-Lys(HEL) ELISA kit

Product Specifications

• Assay principle: Competitive ELISA (detection: 450 nm)
• Specifity: Specific to N-epsilon-Hexanoyl-Lysine adduct
• Measuring range: 2 – 700 nmol/L
• Time to answer: Over night and 2 hours.
• Applications: Urine, serum and cultured cells from human and animals.
• Storage: Store at 2 – 8°C (don’t freeze).
• Expiry: 12 months after the day of manufacturing

References

• Yoji Kato, Yoko Mori, Yuko Makino, Yasujiro Morimitsu, Sadayuki Hiroi, Toshitsugu Ishikawa, Toshihiko Osawa, Formation of N epsilon-(hexanonyl) lysine in protein exposed to lipid hydroperoxide. J Biol Chem 274(29), p20406-20414 (1999)Identification of HEL, which is lysine adduct of 13-HPODE.
• Yoji Kato, Toshihiko Osawa: Detection of lipid hydroperoxide-derived protein modification with polyclonal antibodies. Methods in Molecular Biology, vol. 186, Oxidative stress biomarkers and antioxidant protocols, D Armstrong, Ed., Human Press Inc., NJ, USA, p37-44Immunohistochemical detection of hexanoyl-lysine adduct.
• Kato Y, Miyake Y, Yamamoto K, Shimomura Y, Ochi H, Mori Y, Osawa T, Preparation of a monoclonal antibody to N(epsilon)-(Hexanonyl)lysine: application to the evaluation of protective effects of flavonoid supplementation against exercise-induced oxidative stress in rat skeletal muscle. Biochem Biophys Res Commun 274(2),p389-393(2000)Development and characterization of anti HEL monoclonal antibody.
• Ryo K, Yamada H, Nakagawa Y, Tai Y, Obara K, Inoue H, Mishima K, Saito I, Possible involvement of oxidative stress in salivary gland of patients with Sjogren’s syndrome. Pathobiology 73(5), p252-260 (2006)HEL can be detected in saliva samples from patients with Sjogren’s syndrome.
• Suzuki T, Kazui T, Yamamoto S, Washiyama N, Ohkura K, Ohishi K, Bashar AH, Yamashita K, Terada H, Suzuki K, Akuzawa S, Fujie M, Effect of prophylactically administered edaravone during antegrade cerebral perfusion in a canine model of old cerebral infarction. J Thorac Cardiovasc Surg 133(3),p710-716 (2007)Application to canine serum samples.
• Shimizu K, Ogawa F, Akiyama Y, Muroi E, Yoshizaki A, Iwata Y, Komura K, Bae S, Sato S, Increased Serum Levels of N(epsilon)-(hexanoyl)lysine, A New Marker of Oxidative Stress, in Systemic Sclerosis. J Rheumatol. 2008 Sep 1.Serum HEL concentration from systemic sclerosis is higher than that from healthy subjects.
• Tamura H, Takasaki A, Miwa I, Taniguchi K, Maekawa R, Asada H, Taketani T, Matsuoka A, Yamagata Y, Shimamura K, Morioka H, Ishikawa H, Reiter RJ, Sugino N, Oxidative stress impairs oocyte quality and melatonin protects oocytes from free radical damage and improves fertilization rate. J Pineal Res 44(3),p280-287(2008)Intrafollicular concentration of HEL was significantly reduced by these antioxidant treatment.
• Sakamoto Y, Ishikawa T, Kondo Y, Yamaguchi K, Fujisawa M, The assessment of oxidative stress in infertile patients with varicocele. BJU Int 101(12), p1547-1552 (2008)aAzoospermic and oligospermic patients had a significantly higher HEL concentration in seminal plasma.
• Kageyama Y, Takahashi M, Nagafusa T, Torikai E, Nagano A, Etanercept reduces the oxidative stress marker levels in patients with rheumatoid arthritis. Rheumatol Int 28(3),pp245-251(2008)Urinary HEL level was reduced at 3 and 6 months after the initial treatment with etanercept.
• Rummenie VT, Matsumoto Y, Dogru M, Wang Y, Hu Y, Ward SK, Igarashi A, Wakamatsu T, Ibrahim O, Goto E, Luyten G, Inoue H, Saito I, Shimazaki J, Tsubota K, Tear cytokine and ocular surface alterations following brief passive cigarette smoke exposure. Cytokine 43(2),p200-208(2008)HEL concentration in tear was increased by brief passive exposure to cigarette smoke.

Cholesterol oxidation products, especially 7-Ketocholesterol (7KC), have been the focus of much attention because they are present in human atherosclerotic plaque and display a wide range of atherogenic properties in vitro and to some extant in vivo. This antibody is specific for 7KC. This antibody is useful to detect 7KC in tissue samples. A frozen tissue sample is recommended for immunohistochemistry.

7-Ketocholesterol

Product Information

• 02-1301: Anti 7-ketocholesterol (7-KC) monoclonal antibody (20µg)

Product Specification

• Clone: 35A
• Specificity: Specific for 7-ketocholesterol (7-KC)
• Subclass: Mouse IgG
• Form: Frozen, 100mM PBS containing 0.1% NaN3, 0.5% BSA. IgG is about 1mg/mL
• Applications: Immunohistochemistry (recommended concentration 5 – 10 µg/mL)
• Storage: Store at -20°C

Immunohistochemical detection of 7KC in atherosclerotic aorta. Noriyuki Shibata et al.Tokyo Women’s medical University

References

• Oxysterol mixtures, in atheroma­relevant proportions, display synergistic and proapoptotic effects. David A. Larsson, et. al. Free Radical Biology and Medicine, 41(6), p902­910 (2006)