ADMA (Asymmetric Dimethylarginine) ELISA kit: For Research Use Only Size: 1x96 wells Sensitivity: 0.05 µmol/L Dynamic Range: 0.1 - 5.0 µmol/L Incubation Time: Overnight Sample Type: Serum, Plasma Sample Size: 20 µL Product Developed and Manufactured in Germany
Reference Values (Normal) 0.4 – 0.75 µmol/l (80 – 150 ng/ml) It is suggested that each laboratory establish its own normal ranges
Intended Use The Eagle Biosciences ADMA (Asymmetric Dimethylarginine) ELISA Kit is intended for the quantitative determination of ADMA (Asymmetric Dimethylarginine) in serum or plasma. The ADMA (Asymmetric Dimethylarginine) ELISA Kit is for research use only and not to be used in clinical, therapeutic or diagnostic procedures.
Assay Principle The competitive ADMA (Asymmetric Dimethylarginine) ELISA Kit uses the microtiter plate format. ADMA is bound to the solid phase of the microtiter plate. ADMA in the samples is acylated and competes with solid phase bound ADMA for a fixed number of rabbit anti-ADMA antiserum binding sites. When the system is in equilibrium, free antigen and free antigen-antiserum complexes are removed by washing. The antibody bound to the solid phase ADMA is detected by anti-rabbit/peroxidase. The substrate TMB / peroxidase reaction is monitored at 450 nm. The amount of antibody bound to the solid phase ADMA is inversely proportional to the ADMA concentration of the sample.
Typical Standard Curve:
Assay Background The vascular endothelium plays a central role in the regulation of vascular structure and function, mainly due to the formation of endothelium-derived nitric oxide (NO). NO has been named an “endogenous anti-atherogenic molecule” due to its diverse regulatory functions in vascular homeostasis. NO is formed by the enzyme NO synthetase (NOS) from the amino acid precursor L-arginine. NOS activity can be down-regulated by asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NOS.
The effects of ADMA on NO synthesis and NO-mediated pathophysiological processes have been described in numerous experimental studies. Moreover, elevated ADMA levels in plasma have been found in clinical studies including samples with hypercholesterolemia, hypertension, chronic heart failure, chronic renal failure and other internal disorders. Recent prospective and cross-sectional studies indicated that elevated ADMA levels are a risk factor for future cardiovascular events and total mortality. ADMA may have diagnostic relevance as a novel cardiovascular risk marker.
Cross Reactivity Data: Structural related components were tested for possible interference with the antisera against ADMA used in the ADMA (Asymmetric Dimethylarginine) ELISA kit method. The tested compounds were Arginine, Monomethylarginine (NMMA) und SDMA.
Substance
ED-50-Value (ng/ml)
Cross Reactivity (%)
ADMA
126
100
Arginine
660,000
< 0.02
NMMA
12200
1
SDMA
10500
1.2
Correlation with LC-MS-MS The figure shows the correlation of the ADMA (Asymmetric Dimethylarginine) ELISA kit to the LC-MS-MS method. More data with higher number of samples are evaluated at the moment. Comparative investigations to HPLC without subsequent MS lead to inconsistent results.
Literature Using this ADMA ELISA kit: Schulze F, Wesemann R, Schwedhelm E, Sydow K, Albsmeier J, Cooke JP, Böger RH. Determination of ADMA using a novel ELISA assay. Clin. Chem. Lab. Med. 2004; 42: 1377-1383
Krempl TK, Kähler J, Maas R, Silberhorn L, Meinertz T, Böger RH. Elevation of asymmetric dimethylarginine (ADMA) in patients with unstable angina and recurrent cardiovascular events. Eur. Heart J. 2005; 26: 1846-1851
Schulze F, Maas R, Freese R, Schwedhelm E, Silberhorn L, Böger RH. Determination of a reference value for N,N-dimethyl-L-arginine in 500 subjects. Eur. J. Clin. Invest. 2005; 35 : 622-626
Schnabel R, Blankenberg S, Lubos E, Lackner KJ, Rupprecht HJ, Espinola-Klein C, Jachmann N, Post F, Peetz D, Bickel C, Cambien F, Tiret L, Münzel T. Asymmetric dimethylarginine and the risk of cardiovascular events and death in patients with coronary artery disease: results from the AtheroGene Study. Circ. Res. 2005; 97: e53-59
O`Dwyer MJ, Dempsey F, Crowley V, Kelleher D, McManus R, Ryan T. Septic shock correlates with ADMA levels which may be influenced by a polymorphism in DDAH II: a prospective observational study. Crit. Care 2006; 10: (5): R139
Antoniades C, Tousoulis D, Marinou K, Vasiliadou C, Tentolouris C, Bouras G, Pitsavos C, Stefanidis C. Asymmetrical dimethylarginine regulates endothelial function in methionine-induced but not in chronic homocystinemia in humans: effect of oxidative stress and proinflammatory cytokines Am. J. Clin. Nutr. 2006; 84: 781-788
Wang TZ., Chen WJ., Cheng WC., Lin JW., Chen MF., Lee YT. Relation of improvement in endothelium-dependent flowmediated vasodilation after Rosiglitazone to changes in asymmetric dimethylarginine, endothelin-1, and C-reactive protein in nondiabetic patients with the metabolic syndrome Am. J. Cardiol. 2006; 9: 1057-1062
Wanby P., Nilsson I., Brudin L., Nyhammar I., Gustafsson I., Carlsson M. Increased plasma levels of asymmetric dimethylarginine in patients with carotid stenosis: no evidence for the role of the common FABBP2 A54T gene polymorphism Acta Neurol. Scand. 2007; 115: 90-96
Iribarren C, Husson G, Sydow K, Wang BY, Sidney S, Cooke JP. Asymmetric dimethyl-arginine and coronary artery calcification in young adults entering middle age: the CARDIA Study Eur. J. Cardiovasc. Prev. Rehabil. 2007; 14:222-229 Melikian N, Wheatcroft SB, Ogah OS, Murphy C, Chowienczyk PJ, Wierzbicki AS, Sanders TA, Jiang B, Duncan ER, Shah AM, Kearney MT. Asymmetric dimethylarginine and reduced nitric oxide bioavailability in young Black African men Hypertension 2007; 49: 873-877
Horowitz JD, Heresztyn T. An overview of plasma concentrations of asymmetric dimethylarginine (ADMA) in health and disease and in clinical studies: Methodological considerations. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2007; epub ahead of print
Korish AA, Arafah MM. Catechin combined with vitamins C and E ameliorates insulin resistance (IR) and atherosclerotic changes in aged rats with chronic renal failure (CRF) Arch. Gerontol. Geriatr. 2007; in press
Charitidou C, Farmakiotis D, Zournatzi V, Pidonia I, Pegiou T, Karamanis N, Hatzistilianou M, Katsikis I, Panidis D. The administration of estrogens, combined with anti-androgens, has beneficial effects on the hormonal features and asymmetric dimethyl-arginine levels, in women with the polycystic ovary syndrome Atherosclerosis 2007; in press
