Urea Nitrogen (BUN) detection Kit

Details for Product No. ABIN577679, Supplier: Log in to see
Antigen
Minimum Detection Limit
0.065 mg/dL
Application
Biochemical Assay (BCA)
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Purpose The DetectX® Urea Nitrogen (also called BUN) Detection Kit is designed to quantitatively measureurea nitrogen in a variety of samples.
Brand DetectX®
Sample Type Serum, Plasma, Urine, Saliva, Tissue Culture Supernatant
Detection Method Colorimetric
Specificity Species Independent. Samples Types validated: Serum, Plasma, Urine, Saliva and TCM
Sensitivity 0.30 µg/mL
Characteristics The Urea Nitrogen (also called BUN) Detection Kit is designed to quantitatively measure urea nitrogen in a variety of samples. A urea nitrogen standard calibrated to NIST reference materials is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Samples are mixed with Color Reagents A and B and incubated at room temperature for 30 minutes. The colored product is read at 450 nm. The concentration of urea nitrogen in the sample is calculated, after making a suitable correction for any dilution, using software available with most plate readers. The results are expressed in terms of mg/dL urea nitrogen. If samples are to be expressed in terms of mg/dL urea, the data can be converted using the multiplier 2.14.
Calibated - N-Cal Kit, NIST-Calibrated
Components Clear 96 well Plates - Bags containing 96 well plates 2 or 5 plates
Urea Nitrogen Standard - Urea Nitrogen at 100 mg/dl in a special stabilizing solution. 250 μL or 1 mL Calibrated to NIST Standard Reference Material Lot Number 912a
Color reagent A - An acidic solution of Color reagent A. CAUTION: CAUSTIC 15 mL or 38 mL
Color reagent B - An acidic solution of Color reagent B. CAUTION: CAUSTIC 15 mL or 38 mL
Material not included Distilled or deionized water free of urea. 96 well plate reader capable of reading optical absorption at 450 nm.
Software for converting optical density (OD) readings from the plate reader and carrying out four parameter logistic curve (4PLC) fitting.
Background Urea is a by-product of protein metabolism by the liver, and is therefore removed from the blood by the kidneys. Urea freely filters through the glomerulous, but is reabsorbed by the renal tubules in a flow-dependent fashion. The higher the flow rate, the greater amount of urea nitrogen is cleared from circulation and eliminated through the kidneys. As a result, the level of circulating urea nitrogen, along with serum creatinine, serves as a primary measure of kidney function. Normal adult Blood Urea Nitrogen (BUN) levels should be between 7 and 21 mg urea nitrogen per 100 mL blood (mg/dL)1. Azotemia, poor kidney function, will cause elevated BUN levels (≥ 50 mg/dL) and is associated with acute kidney failure or injury, severe acute pancreatitis, congestive heart failure or gastrointestinal bleeding2-5. Azotemia also can occur with dehydration, as a result of alcohol abuse, or high protein diets. Lower than expected BUN levels are usually not clinically predictive, but are primarily associated with liver disease or malnutrition, including malabsorption and low protein diets6. Urine and saliva are considered to be acceptable non-invasive samples for measurement of urea nitrogen7. Serum creatinine is another metabolic waste product freely filtered by the glumerulous, but does not undergo tubular reabsorption. Its steady rate of elimination is frequently used to generate an index or ratio with BUN values for normalized evaluations. Easy to use Serum Creatinine and Urinary Creatinine Detection kits are also available from The Supplier (see Related Products)
Application Notes Urea nitrogen is identical across all species and this kit will measure urea nitrogen from sources other than human.
The end user should evaluate recoveries of urea nitrogen in samples from other species being tested.
The kit will measure urea nitrogen in low concentration samples such as RPMI cell culture media, however the media should not contain Phenol Red.
If samples need to be stored after collection, we recommend storing them at -70 °C or lower, preferably after being frozen in liquid nitrogen.
This assay has been validated for serum, plasma and urine.
Samples containing visible particulate should be centrifuged prior to using.
Comment

Sample values: Six random adult human serum and plasma samples were diluted and tested in the assay.
The serum samples ranged from 15.6 to 22.3 mg/dL with an average of 18.6 mg/dL BUN while EDTA and heparin plasma samples ranged from 13.6 to 23.7 mg/dL with an average BUN of 18.1 mg/dL.
Six random saliva samples were clarified, diluted and tested in the kit.
The Urea Nitrogen values ranged from 4.3 to 11.9 mg/dL, with an average concentration of 8.7 mg/dL.
Six random urines were also diluted and tested in the kit.
The Urea Nitrogen values widely ranged from 37.2 to 1007.2 mg/dL as expected for random urine sampling.

Assay Time 1 h
Plate 96 wells
Protocol A urea nitrogen standard calibrated to NIST reference materials is provided to generate a standard curve for the assay and all samples should be read off the standard curve.
Samples are mixed with Color Reagents A and B and incubated at room temperature for 30 minutes.
The colored product is read at 450 nm.
The concentration of urea nitrogen in the sample is calculated, after making a suitable correction for any dilution, using software available with most plate readers.
The results are expressed in terms of mg/dL urea nitrogen.
If samples are to be expressed in terms of mg/dL urea, the data can be converted using the multiplier 2.14.
Sample Preparation

Dilute sample with distilled or deionized water prior to running in the assay. For serum or plasma, the recommended dilution is ≥ 1:10 and ≥1:20 respectively. Saliva should be clarified by freeze/ thawing, followed by centrifugation at 14,000 rpm at 4 °C for 10 minutes. The saliva supernatant should be diluted at least 1:2 before measuring in the assay. For urine, where concentrations of urea are higher, the recommended final dilution is ≥ 1:100. For highly colored samples, dilution greater than 1:10 or 1:100 may be necessary.

Assay Procedure

Use the plate layout sheet on the back page to aid in proper sample and standard identification.
1. Pipet 50 μL of samples or appropriate standards into duplicate wells in the plate.
2. Pipet 50 μL of water into duplicate wells as the Zero standard.
3. Add 75 μL of Color Reagent A to each well using a repeater pipet.
4. Add 75 μL of Color Reagent B to each well using a repeater pipet.
5. Incubate at room temperature for 30 minutes.
6. Read the optical density at 450 nm. ® 6 EXPECT ASSAY ARTISTRY

Calculation of Results

Average the duplicate OD readings for each standard and sample.
Create a standard curve by reducing the data using computer software capable of generating a four-parameter logistic curve (4PLC) fit, after subtracting the mean OD's for the blank.
The sample concentrations obtained should be multiplied by the dilution factor to obtain neat sample values.

Restrictions For Research Use only
Precaution of Use As with all such products, this kit should only be used by qualified personnel who have had laboratory safety instruction.
The complete insert should be read and understood before attempting to use the product. the Color reagents A and B are both strong acid solutions and should be handled like any laboratory acid.
Storage 4 °C,RT
Storage Comment All components of this kit should be stored at room temperature until the expiration date of the kit.
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