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Uric Acid Assay Kit

Uric acid is an end product of purine nucleotide metabolism in primate mammals, and is mainly excreted in urine. Serum uric acid concentrations are dependent upon the rate of uric acid produced and renal uric acid excretion. Hyperuricemia (elevated level of serum uric acid) induces or facilitates gout, kidney stones, metabolic syndrome, hypertension, and renal and cardiovascular disease Moreover, the serum UA level is considered to be a marker of oxidative stress. Eton’s Uric acid assay kit provides a reliable, simple, rapid, and suitable for either manual or automated procedures for quantifying uric acid in biological samples such as blood (serum and plasma), urine, etc. In the assay, uric acid is oxidized by enzyme reactions to yield color product, which can be measured at 570 nm for colorimetric assay, and the color intensity is proportional to uric acid contents, therefore the sample uric acid concentration can be accurately calculated based on the uric acid curve.

Sku #Product NameProduct SizePriceQTY
120006100P Uric Acid Assay Kit 100 Assays w/Reader Plate(1x) 100 Assays w/Reader Plate(1x) $150.00 USD
1200061002 Uric Acid Assay Kit 100 assays 100 assays $130.00 USD
 

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Sample Typle

Plasma,Serum,Cell culture supernatant, other body fluid

Components

Method

Colorimetric method at 570nm

Standard Curve

Sensitivity

10µM-500µM

Reaction Time

30 minutes

Applications

For biological research:
Uric Acid measurement in biological samples
For drug/pharm research:
Drug influence on purine metabolism

Storage

-80°C

Notes

Material needed but not supplied

A plate reader capable of measuring absorbance at 570nm

Adjustable pipettes and a repeat pipettor

The distilled water(milliQ or HPLC-grade)

Clear Flat bottom 96-well plates if not included in the kit purchased

Shipping Icepacks

Reagent Preparation

Note: All reagents are frozen. We recommend you spin small vials before opening.

1. Uric Acid Assay Buffer(10x)
Please do not dilute in the provided Uric Acid Assay Buffer bottle itself.

The bottle contains 10ml of 10xUric Acid Assay Buffer. The Assay Buffer must be equilibrated to room temperature before use. Please make appropriate amount of1xUric Acid Assay Buffer for the assay, for example, by diluting 1ml of 10x Uric Acid Assay Buffer with 9 ml of dH2O. Prepare the additional diluted assay buffer if needed. Store at -80?C.

2. Uric Acid Standard (2x)

Please do not dilute in the provided Uric Acid Standard vial itself.

The vial contains 500µl of 1mM Uric Acid Standard. The standard must be equilibrated to room temperature before use. Dilute 500µl of 1mM Uric Acid Standard with 500µl of diluted Uric Acid Assay buffer to prepare 500µM of Uric Acid Standard. 1ml of 500µM Uric Acid Standard is enough for making 3 standard curves if assayed in duplicate.Store at -80?C.

3. Assay Probe
The vial contains 45μl of Assay Probe. It is sufficient for 100 wells. The assay probe must be equilibrated to room temperature before use.Protect from light. Store at -80?C.

4. Uric Acid Assay Enzyme Mix
The vial contains 30μl of Uric Acid Assay Enzyme Mix. The enzyme mix must be thawed on ice before use. It is sufficient for 100 wells. Store at -80?C.

Protocol

1.Sample Preparation

Blood (serum/plasma), urine, other body fluids can be measured directly by a series of dilutions of the sample (1/2; 1/4; 1/8; …) to ensure the readings are within the standard curve range.Your samples can be diluted with dH2O or diluted Uric Acid Assay Buffer.

Add 50μl of samples to each well.
We recommend that samples be assayed in duplicate or triplicate.


Note:
If prepared samples are not assayed the same day, store the samples at -80?C. The samples will be stable for one month while stored at -80?C. For frozen samples, dilutions of samples must be done right before assaying.

2.Standard Curve Prepation

Add 50μl, 40μl, 30μl, 20μl, 10μl, 5μl, 1μl, and 0μl of diluted Uric Acid Standard to each well, then adjust volume to 50μl/well with diluted Uric Acid Assay Buffer.
We recommend that Uric Acid Standards be assayed in duplicate. A standard curve has to be run in each assay.

3.Reaction Solution Preparation
50μl Reaction Mix is required for each well and 1ml of Reaction Solution is prepared with the followings:
984µl Uric Acid Assay Buffer (1x)
8 µl Uric Acid Assay Enzyme Mix
8 µl Assay Probe

1ml of Reaction Solution is enough for ~20 wells.

Prepare appropriate amount of Reaction Solution for the number of assays to be performed.

Note:Please do not keep any leftover Reaction Solution as it has to be made fresh right beforeyou run assays. If old Reaction Solution is used, assays will not work.

4.Perform the assay

a. Mix the reaction solution well and add 50μl of the reaction solution to each well containing the Uric Acid standards and test samples.

b. Incubate the reaction for 30 minutes in a 37?C incubator.

c. Measure absorbance at 570 nm in a microplate reader.

5. Calculation

a. Average the OD570 nm values of replicate wells of each Uric Acid Standard, test samples, and blank. Then, correct background by subtracting the value derived from the blank from all sample readings.

b. Make a standard curve by plotting OD570 nm values from each Uric Acid Standards as a function of Uric Acid concentration. This can be done with excel spreadsheet.

c. Calculate the value of Uric Acid in samples using the equation obtained from the linear regression of the standard curve.

Uric Acid(μM)= [ (Corrected absorbance)-(y-intercept)]/Slope


Material Safety Data Sheet

Date:Oct 26,2015

1. PRODUCT AND COMPANY IDENTIFICATION

1.1 Product identifier

Product name: Uric Acid Assay Kit

Catalog No: 1200061002,120006100P

1.2 Relevant identified uses of the substance or mixture and uses advised against

For research use only.

1.3 Details of the supplier of the safety data sheet

Company:Eton Bioscience Inc

TollFree:1-800-758-1630

Tel:1-800-758-1630

Fax:1-800-507-2912

1.4 Emergency telephone number

1-800-758-1630

2. HAZARDS IDENTIFICATION

2.1 Classification of the substance or mixture

Not a hazardous substance or mixture

2.2 GHS Label elements, including precautionary statements

Not a hazardous substance or mixture

2.3 Other hazards

None

3. COMPOSITION/INFORMATION ON INGREDIENTS

3.1 Substances

Uric Acid Standard

Uric Acid Assay Enzyme Mix

Components

CAS#

Components

CAS#

Uric Acid

69-93-2

Uricase

9002-12-4

Lithium Carbonate

554-13-2

Tris-HCl pH7.5

N/A

Water

7732-18-5

Water

7732-18-5

Uric Acid Assay Buffer

Assay Probe

Components

CAS#

Components

CAS#

Tris-HCl pH7.5

N/A

DMSO

67-68-5

Water

7732-18-5













4. FIRST AID MEASURES

4.1 Description of first aid measures

Eye contact

Check for and remove contact lenses and flush with copious amounts of water; assure adequate flushing by separating the eyelids with fingers; call a physician

Skin Contact

Flush with copious amounts of water; remove contaminated clothing and shoes; call a physician

Inhalation

Remove to fresh air. If not breathing give artificial respiration ;call a physician

Ingestion

If swallowed, never give anything by mouth to an unconscious person. Rinse mouth with water ;call a physician

4.2 Most important symptoms and effects, both acute and delayed

No information available

4.3 Indication of any immediate medical attention and special treatment needed.

No information available

5. FIRE FIGHTING MEASURES

5.1 Extinguishing media

Suitable extinguishing media

Water spray, alcohol-resistant foam, dry chemical or carbon dioxide

5.2 Special hazards arising from the substance or mixture

No information available

5.3 Advice for firefighters

Wear self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes.

6. ACCIDENTAL RELEASE MEASURES

6.1 Personal precautions, protective equipment and emergency procedures

Use full personal protective equipment. Evacuate personnel to safe areas. Avoid breathing vapors,mist or gas.

6.2 Environmental precautions

Prevent further leakage or spillage. Prevent product from entering drain.

6.3 Methods and material for containment and cleaning up

Keep in suitable, closed containers for disposal

7. HANDLING AND STORAGE

7.1 Precautions for safe handling

Avoid skin/eye contact. Use protective equipment as needed. Wash contaminated clothing before reuse.

7.2 Conditions for safe storage,including any incompatibilities

Keep container tightly closed. Store at -80?C.

7.3 Specific end use(s)

No data available.

8. EXPOSURE CONTROLS/PERSONAL INFORMATION

8.1 Control Parameters

This product contains no hazardous materials with occupational exposure limits.

8.2 Exposure controls

Engineering controls

Provide shower and eye wash station

Personal protective equipment

Eye Protection

Wear safety goggles

Skin Protection

Wear protective clothing

Hand Protection

Wear protective gloves

Respiratory protection

Wear NIOSH/MSHA approved respirators

9. PHYSICAL AND CHEMICAL PROPERTIES

9.1 Information on basic physical and chemical properties

Uric Acid Assay Buffer

Appearance

Liquid, colorless

pH

7.5

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

Uric Acid Standard

Appearance

Liquid, colorless

pH

No data available

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

Uric Acid Assay Enzyme Mix

Appearance

Liquid, colorless

pH

No data available

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

Assay Probe

Appearance

Liquid, colorless

pH

No data available

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

9.2 Other safety information

No data available.

10. STABILITY AND REACTIVITY

10.1Reactivity

No data available

10.2Chemical stability

Stable under recommended storage conditions.

10.3Possibility of hazardous reactions

No data available.

10.4Conditions to avoid

No data available

10.5Incompatible materials

Strong oxidizing agents

10.6Hazardous decomposition products

Hazardous decomposition products formed under fire conditions: carbon oxides

11. TOXICOLOGICAL INFORMATION

11.1Information on toxicological effects

Toxicological effects on this product have not been thoroughly studied.

12. ECOLOGICAL INFORMATION

12.1Toxicity

Avoid release into environment

12.2Persistence and degradability

No data available

12.3.Bioaccumulative potential

No data available

12.4Mobility in soil

No data available

12.5Results of PBT and vPvB assessment

No data available

12.6Other adverse effects

No data available

13. DISPOSAL INFORMATION

13.1Waste treatment methods

Dispose in accordance with local, state, and federal regulations.

14. TRANSPORT INFORMATION

DOT:

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport
IATA:

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport

ADR/RID

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport

15. REGULATORY INFORMATION

EU Risk and Safety phrases:

R36/37/38: irritating to eyes/respiratory system/skin

16. OTHER INFORMATION

The information above is believed to be accurate and represents the best information currently available to us. However, we make no warranty of merchantability or any other warranty, express or implied, with respect to such information. Eton Bioscience Inc. shall not be held liable for any damages or other consequences resulting from handling or from contact with the above product.


1. Milner JA, Perkins EG., Determination of uric acid in biological fluids by high performance liquid chromatography. Anal Biochem, 88:560–5 (1978).
2. Pachla LA, Kissinger PT. Measurement of serum uric acid by liquid chromatography. Clin Chem, 25:1847–52 (1979).
3. Watanabe, S., Kang, D.-H., Feng, L., et al. Uric acid, hominoid evolution, and the pathogenesis of salt-sensitivity. Hypertension 40, 355-360 (2002).
4. Arias-Mendoza F, Piña E. (1991). A sensitive multienzymatic assay for the measurement of Uric Acid, dihydroxyacetone phosphate, oxaloacetate, and acetoacetate in clear extracts from biological samples. Prep Biochem. 21(4):211-4.
5. Chariot P. et al (1994). Optimal handling of blood samples for routine measurement of lactate and Uric Acid. Arch Pathol Lab Med. 118(7):695-7.

Esquivel, Carlos J., Bryan J. Cassone, and Peter M. Piermarini. “A de Novo Transcriptome of the Malpighian Tubules in Non-Blood-Fed and Blood-Fed Asian Tiger Mosquitoes Aedes Albopictus: Insights into Diuresis, Detoxification, and Blood Meal Processing.” Ed. Stephen Tobe. PeerJ 4 (2016): e1784. PMC. Web. 9 Aug. 2017.

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Eton Bioscience, Inc.
5820 Oberlin Drive, Suite 108
San Diego, CA 92121
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