ELISA Kit for Asymmetrical Dimethylarginine (ADMA)

N,N-Dimethylarginine; NG,NG-Dimethylarginine dihydrochloride; Asymmetric Dimethylarginine

Specificity

This assay has high sensitivity and excellent specificity for detection of Asymmetrical Dimethylarginine (ADMA).
No significant cross-reactivity or interference between Asymmetrical Dimethylarginine (ADMA) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Asymmetrical Dimethylarginine (ADMA) and the recovery rates were calculated by comparing the measured value to the expected amount of Asymmetrical Dimethylarginine (ADMA) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-92 87
EDTA plasma(n=5) 92-101 96
heparin plasma(n=5) 96-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Asymmetrical Dimethylarginine (ADMA) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Asymmetrical Dimethylarginine (ADMA) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Asymmetrical Dimethylarginine (ADMA) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-101% 85-97% 98-105% 93-101%
EDTA plasma(n=5) 86-96% 84-101% 91-98% 95-102%
heparin plasma(n=5) 85-102% 98-105% 83-92% 90-101%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
PLoS One. Interactions among Vascular-Tone Modulators Contribute to High Altitude Pulmonary Edema and Augmented Vasoreactivity in Highlanders PubMed: PMC3439466
Cytokine Effect of CCL5 on dimethylarginine dimethylaminohydrolase-1 production in vascular smooth muscle cells from spontaneously hypertensive rats. Pubmed: 23834755
Neurol Res. Effect of 18β-glycyrrhetinic acid on cerebral vasospasm caused by asymmetric dimethylarginine after experimental subarachnoid hemorrhage in rats. Pubmed:25475507
Int J Clin Exp Med. Correlation research on ADMA plasma levels and left ventricular function of peritoneal dialysis patients Pubmed:25550968
Clin Respir J Asymmetric dimethylarginine contributes to airway nitric oxide deficiency in patients with COPD PubMed: 26076870
Cytokine Dimethylarginine dimethylaminohydrolase-1 mediates inhibitory effect of interleukin-10 on angiotensin II-induced hypertensive effects in vascular smooth muscle cells of spontaneously hypertensive rats PubMed: 26375520
Neurological Research Effect of 18β-glycyrrhetinic acid on cerebral vasospasm caused by asymmetric dimethylarginine after experimental subarachnoid hemorrhage in rats PubMed: 25475507
Scientific Reports Dimethylarginine Dimethylaminohydrolase 2 (DDAH 2) Gene Polymorphism, Asymmetric Dimethylarginine (ADMA) Concentrations, and Risk of Coronary Artery Disease: A Case-Control Study pubmed:27677852
clinica chimica acta Increased serum concentrations of asymmetric dimethylarginine (ADMA) in patients with early-onset coronary artery disease. pubmed:27884754
The Journal of Physiology Asymmetric dimethylarginine (ADMA) elevation and arginase up-regulation contribute toendothelial dysfunction related to insulin resistance in rats and morbidly obese humans. pubmed:26840628
Angiogenesis Dimethylarginine dimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostate cancer, and its overexpression conveys tumor growth and angiogenesis by metabolizing asymmetric dimethylarginine (ADMA). pubmed:29150732
Scientific Reports A Novel Role of Irbesartan in Gastroprotection against Indomethacin-Induced Gastric Injury in Rats: Targeting DDAH/ADMA and EGFR/ERK Signaling Pubmed:29523851
Mechanisms of Ageing and Development Multivessel analysis of progressive vascular aging in the rat: Asynchronous vulnerability among vascular territories Pubmed:29605639
Nutrition, Metabolism and Cardiovascular Diseases Arginine bioavailability and endothelin-1 system in the regulation of vascular function of umbilical vein endothelial cells from intrauterine growth restricted newborns Doi: 10.1016/j.numecd.2018.09.002
Biochemical and Biophysical Research Communications Probucol improves erectile function via Activation of Nrf2 and coordinates the HO-1/DDAH/PPAR-γ/eNOS pathways in streptozotocin-induced diabetic rats Pubmed: 30454888
Brazilian Journal of Pharmaceutical Science Demographic, clinical and lifestyle predictors for severity of erectile dysfunction and biomarkers level in Malaysian patients
ACS Combinatorial Science Novel cellularly active inhibitor regresses DDAH1 induced prostate tumor growth by restraining tumor angiogenesis through targeting DDAH1/ADMA/NOS pathway Pubmed: 30673277
International Journal of the Cardiovascular Academy Nebivolol prevents the increase of asymmetric dimethylarginine and oxidants in hyperhomocysteinemic rats
Oxid Med Cell Longev The Role of Reactive Oxygen Species and Nitric Oxide in the Inhibition of Trichophyton rubrum Growth by HaCaT Cells Pubmed: 32104540
Molecular Medicine Reports Blueberry anthocyanin‑enriched extract ameliorates transverse aortic constriction‑induced myocardial dysfunction via the DDAH1/ADMA/NO signaling pathway in … Pubmed: 31746378
International Journal of Nephrology Effect of a Supervised Peridialytic Exercise Program on Serum Asymmetric Dimethylarginine in Maintenance Hemodialysis Patients Pubmed: 33163233
Journal of Clinical Medicine The Usefulness of Serum Biomarkers in the Early Stages of Diabetic Retinopathy: Results of the EUROCONDOR Clinical Trial Pubmed: 32344735
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub Levels of endothelial substances in patients with newly identified hypertension compared with healthy controls 33463628
WIENER KLINISCHE WOCHENSCHRIFT Non-functioning adrenal incidentalomas may increase toxic metabolites 34255169
pharmaceuticals Effect of a Low Dose of Carvedilol on Cyclophosphamide-Induced Urinary Toxicity in Rats—A Comparison with Mesna 34959638
Int J Biochem Cell Biol HIF-1α and Nrf2 regulates hypoxia induced overexpression of DDAH1 through promoter activation in prostate cancer Pubmed:35644470
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