Uric Acid Assay Kit

Catalog No. ABIN1000270

Product Details

Target: Uric Acid

Application: Biochemical Assay (BCA)

Sample Type: Plasma, Serum, Urine

Specificity: 0.2 mg/dL (13 μM)

Characteristics: Sensitive and accurate. Use 5 µL samples. Linear detection range 0.22 mg/dL (13µM) to 30 mg/dL (2380µM) uric acid in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 30 min. Can be readily automated as a high-throughput assay in 96-well plates for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Cuvet or 96-well plate assay.
Low interference in biological samples.
No pretreatments are needed. Assays can be directly performed on serum samples.

Components: Reagent A: 50 mL. Reagent B: 6 mL. Reagent C: 6 mL. Standard: 1 mL 10 mg/dL uric acid. Blank Control: 1 mL.

Material not included: Pipeting devices and accessories (e.g. 5 µL). Clear bottom 96-well plates (e.g. Corning Costar). 96-well plate absorbance (590nm) reader. Cuvets for measuring optical density at 510-630nm. Spectrophotometer for measuring absorbance at 590nm.

Application Details

Application Notes: Direct Assays: uric acid in serum, plasma, urine and other biological samples.
Drug Discovery/Pharmacology: effects of drugs on uric acid metabolism.

Protocol: Procedure using 96-well plate:
1. Set up standards and samples. Transfer 5 µL Blank, Standard and samples in duplicate wells of a clear bottom 96-well plate.
2. Add 200 µL working reagent and tap lightly to mix.
3. Incubate 30 min at room temperature and read optical density at 510- 630nm (peak absorbance at 590nm).

Procedure using cuvette:
1. Set up test tubes labeled Blank, Standard, Samples. Transfer 20 µL Blank, Standard and samples to appropriately labeled tubes.
2. Add 1000 µL working reagent and tap lightly to mix.
3. Incubate 30 min at room temperature and read optical density at 590nm (510nm-630nm).

Reagent Preparation:

Shake Reagent C before use. Prepare enough working reagent by mixing 10 volumes of Reagent A, 1 volume Reagent B and 1 volume Reagent C. Fresh reconstitution is recommended. Equilibrate to room temperature before assay. Metal chelators (e.g. EDTA) interfere with this assay and should be avoided.

Calculation of Results:

Normal serum uric acid values: 1.0 to 7.0 mg/dL.
Conversions: 1 mg/dL uric acid equals 59.5 µM, 0.001% or 10 ppm.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Karaouzene, Merzouk, Aribi, Merzouk, Berrouiguet, Tessier, Narce: "Effects of the association of aging and obesity on lipids, lipoproteins and oxidative stress biomarkers: a comparison of older with young men." in: Nutrition, metabolism, and cardiovascular diseases : NMCD, Vol. 21, Issue 10, pp. 792-9, (2011) (PubMed).

Smith, Toomey, Walker, Braun, Wolf, McGraw, Sweazea: "Naturally high plasma glucose levels in mourning doves (Zenaida macroura) do not lead to high levels of reactive oxygen species in the vasculature." in: Zoology (Jena, Germany), Vol. 114, Issue 3, pp. 171-6, (2011) (PubMed).

Xu, Rao, Groh, Spies, Gattuso, Kaufman, Plate, Prinz: "Major histocompatibility complex class I-related chain A/B (MICA/B) expression in tumor tissue and serum of pancreatic cancer: role of uric acid accumulation in gemcitabine-induced MICA/B expression." in: BMC cancer, Vol. 11, pp. 194, (2011) (PubMed).

Belarbi, Bendimerad, Sour, Soualem, Baghdad, Hmimed, Chemat, Visioli: "Oleaster oil positively modulates plasma lipids in humans." in: Journal of agricultural and food chemistry, Vol. 59, Issue 16, pp. 8667-9, (2011) (PubMed).

Moon, Tae, Kim, Gyu Jeon, Oh, Song Gho, Zhu, Kim: "Conversion of Th17-type into Th2-type inflammation by acetyl salicylic acid via the adenosine and uric acid pathway in the lung." in: Allergy, Vol. 65, Issue 9, pp. 1093-103, (2010) (PubMed).

Yu, Sánchez-Lozada, Johnson, Kang: "Oxidative stress with an activation of the renin-angiotensin system in human vascular endothelial cells as a novel mechanism of uric acid-induced endothelial dysfunction." in: Journal of hypertension, Vol. 28, Issue 6, pp. 1234-42, (2010) (PubMed).

Kono, Chen, Ontiveros, Rock: "Uric acid promotes an acute inflammatory response to sterile cell death in mice." in: The Journal of clinical investigation, Vol. 120, Issue 6, pp. 1939-49, (2010) (PubMed).

Whidden, McClung, Falk, Hudson, Smuder, Nelson, Powers: "Xanthine oxidase contributes to mechanical ventilation-induced diaphragmatic oxidative stress and contractile dysfunction." in: Journal of applied physiology (Bethesda, Md. : 1985), Vol. 106, Issue 2, pp. 385-94, (2009) (PubMed).

DiSilvestro, DiSilvestro, DiSilvestro: "Pomegranate extract mouth rinsing effects on saliva measures relevant to gingivitis risk." in: Phytotherapy research : PTR, Vol. 23, Issue 8, pp. 1123-7, (2009) (PubMed).

Brosnahan, Mantz, Squier, Peterson, Schlievert: "Cytolysins augment superantigen penetration of stratified mucosa." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 182, Issue 4, pp. 2364-73, (2009) (PubMed).

Viel, Benkirane, Javeshghani, Touyz, Schiffrin: "Xanthine oxidase and mitochondria contribute to vascular superoxide anion generation in DOCA-salt hypertensive rats." in: American journal of physiology. Heart and circulatory physiology, Vol. 295, Issue 1, pp. H281-8, (2008) (PubMed).

Kaya, Koc, Sogut, Duru, Yilmaz, Uz, Durgut: "The protective effect of N-acetylcysteine against cyclosporine A-induced hepatotoxicity in rats." in: Journal of applied toxicology : JAT, Vol. 28, Issue 1, pp. 15-20, (2007) (PubMed).

Kamel: "Conventional and planar chip sensors for potentiometric assay of uric acid in biological fluids using flow injection analysis." in: Journal of pharmaceutical and biomedical analysis, Vol. 45, Issue 2, pp. 341-8, (2007) (PubMed).

Thangaraju, Ananth, Martin, Roon, Smith, Sterneck, Prasad, Ganapathy: "c/ebpdelta Null mouse as a model for the double knock-out of slc5a8 and slc5a12 in kidney." in: The Journal of biological chemistry, Vol. 281, Issue 37, pp. 26769-73, (2006) (PubMed).

Target Details

Target: Uric Acid

Target Type: Chemical

Background: Quantitative determination of uric acid by colorimetric (590nm) method.
Procedure: 30 min.

Uric acid is the waste product produced from the degradation of purines. In healthy human, uric acid is filtered and removed from the blood by the kidneys and excreted into urine. Because a number of kidney diseases are known to affect uric acid levels, uric acid determination is thus important and useful in diagnosing and evaluating kidney diseases. For example, when uric acid is present in the blood at abnormally high levels, it tends to crystallize in body joints, resulting in gout, a very painful inflammatory condition. Increased levels of uric acid are also known to be associated with uremia, leukemia and pneumonia. Simple, direct and automation-ready procedures for measuring uric acid concentration in blood are becoming popular in Research and Drug Discovery. This uric acid assay kit is designed to measure uric acid directly in serum without any pretreatment. The improved method utilizes 2,4,6-tripyridyl-s-triazine that forms a blue colored complex specifically with iron in the presence of uric acid. The intensity of the color, measured at 590nm, is directly proportional to the uric acid concentration in the serum. The optimized formulation substantially reduces interference by substances in the raw samples.