Urea Nitrogen (BUN) detection Kit

Catalog No. ABIN577679

Product Details

Target: Urea Nitrogen (BUN)

Detection Method: Colorimetric

Minimum Detection Limit: 0.065 mg/dL

Application: Biochemical Assay (BCA)

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: Cell Culture Supernatant, Plasma, Saliva, Serum, Urine

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.

Application Details

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

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

Handling

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.

References

Choi, Lee, HwangBo, Kwon, Kim, Ji, Hong, Kim, Park, Hwang, Moon, Yun, Kim, Choi: "Citrus unshiu peel suppress the metastatic potential of murine melanoma B16F10 cells in vitro and in vivo." in: Phytotherapy research : PTR, Vol. 33, Issue 12, pp. 3228-3241, (2020) (PubMed).

Li, Hu, Li, Liang, Xiang, Hsiao, Nguyen, Park, Egranov, Ambati, Putluri, Hawke, Han, Hung, Danesh, Yang, Lin: "PTEN-induced partial epithelial-mesenchymal transition drives diabetic kidney disease." in: The Journal of clinical investigation, Vol. 129, Issue 3, pp. 1129-1151, (2019) (PubMed).

Zhang, Bush, Yan, Chen: "Gemcitabine nanoparticles promote antitumor immunity against melanoma." in: Biomaterials, Vol. 189, pp. 48-59, (2019) (PubMed).

Yu, Chun, Ha, Kim, Lih, Kim, Kim, Chung, Song, Yoo, Chung, Han, Kim, Kwon: "In Vivo Safety and Regeneration of Long-Term Transported Amniotic Fluid Stem Cells for Renal Regeneration." in: Tissue engineering and regenerative medicine, Vol. 16, Issue 1, pp. 81-92, (2019) (PubMed).

Yan, Deng, Zhao, Ye, Fang, Meng, Wang, Luo, Liu, Li: "Establishment and characterization of an immortalized human hepatocyte line for the development of bioartificial liver system." in: Cytotechnology, (2018) (PubMed).

Kaneko, Cho, Sato, Goto, Yamamoto, Goto, Madaio, Narita: "Attenuated Macrophage Infiltration in Glomeruli of Aged Mice Resulting in Ameliorated Kidney Injury in Nephrotoxic Serum Nephritis." in: The journals of gerontology. Series A, Biological sciences and medical sciences, (2018) (PubMed).

Sato, Saigusa, Mishima, Uchida, Miura, Morikawa-Ichinose, Kisu, Sekimoto, Saito, Oe, Matsumoto, Tomioka, Mori, Takahashi, Sato, Abe, Niwa, Ito: "Impact of the Oral Adsorbent AST-120 on Organ-Specific Accumulation of Uremic Toxins: LC-MS/MS and MS Imaging Techniques." in: Toxins, Vol. 10, Issue 1, (2018) (PubMed).

Ara, Fadriquela, Ahmed, Bajgai, Sajo, Lee, Kim, Jung, Kim, Kim, Shim, Lee: "Hydrogen Water Drinking Exerts Antifatigue Effects in Chronic Forced Swimming Mice via Antioxidative and Anti-Inflammatory Activities." in: BioMed research international, Vol. 2018, pp. 2571269, (2018) (PubMed).

Kishimoto, Kirino, Tamura, Takeno, Kunishita, Takase-Minegishi, Nakano, Kato, Nagahama, Yoshimi, Igarashi, Aoki, Nakajima: "Dysregulated heme oxygenase-1low M2-like macrophages augment lupus nephritis via Bach1 induced by type I interferons." in: Arthritis research & therapy, Vol. 20, Issue 1, pp. 64, (2018) (PubMed).

Chen, Wang, Jiang, Zhao, Fan, Wang, Peng: "Huangqi (astragalus) decoction ameliorates diabetic nephropathy via IRS1-PI3K-GLUT signaling pathway." in: American journal of translational research, Vol. 10, Issue 8, pp. 2491-2501, (2018) (PubMed).

Kovalčíková, Janšáková, Gyurászová, Podracká, Šebeková, Celec, Tóthová: "Salivary creatinine and urea are higher in an experimental model of acute but not chronic renal disease." in: PLoS ONE, Vol. 13, Issue 7, pp. e0200391, (2018) (PubMed).

Burgess, Hunt, Kraus, Rolland: "Quantifying hormones in exhaled breath for physiological assessment of large whales at sea." in: Scientific reports, Vol. 8, Issue 1, pp. 10031, (2018) (PubMed).

Ranghino, Bruno, Bussolati, Moggio, Dimuccio, Tapparo, Biancone, Gontero, Frea, Camussi: "The effects of glomerular and tubular renal progenitors and derived extracellular vesicles on recovery from acute kidney injury." in: Stem cell research & therapy, Vol. 8, Issue 1, pp. 24, (2017) (PubMed).

Guo, Cao, Chu, Wang, Zang, Mao, Wang, Wang, Liu, Zhang, Peng: "Astragaloside IV Attenuates Podocyte Apoptosis Mediated by Endoplasmic Reticulum Stress through Upregulating Sarco/Endoplasmic Reticulum Ca(2+)-ATPase 2 Expression in Diabetic Nephropathy." in: Frontiers in pharmacology, Vol. 7, pp. 500, (2017) (PubMed).

Weaver, Prichard, Maerz, Prichard, Endres, Hernández-Castellano, Akins, Bruckmaier, Hernandez: "Elevating serotonin pre-partum alters the Holstein dairy cow hepatic adaptation to lactation." in: PLoS ONE, Vol. 12, Issue 9, pp. e0184939, (2017) (PubMed).

Call, Donet, Martin, Sharma, Chen, Zhang, Cai, Galarreta, Okutsu, Du, Lira, Zhang, Mehrad, Annex, Klibanov, Bowler, Laubach, Peirce, Yan: "Muscle-derived extracellular superoxide dismutase inhibits endothelial activation and protects against multiple organ dysfunction syndrome in mice." in: Free radical biology & medicine, Vol. 113, pp. 212-223, (2017) (PubMed).

Burdick Sanchez, Carroll, Broadway, Hughes, Roberts, Richeson, Schmidt, Vann: "Cattle temperament influences metabolism: metabolic response to glucose tolerance and insulin sensitivity tests in beef steers." in: Domestic animal endocrinology, Vol. 56, pp. 85-95, (2016) (PubMed).

Soni, Adebiyi: "Early septic insult in neonatal pigs increases serum and urinary soluble Fas ligand and decreases kidney function without inducing significant renal apoptosis." in: Renal failure, pp. 1-12, (2016) (PubMed).

Buntyn, Burdick Sanchez, Schmidt, Erickson, Sieren, Jones, Carroll: "The metabolic, stress axis, and hematology response of zilpaterol hydrochloride supplemented beef heifers when exposed to a dual corticotropin-releasing hormone and vasopressin challenge." in: Journal of animal science, Vol. 94, Issue 7, pp. 2798-810, (2016) (PubMed).

Hayashi, Oe, Fushima, Sato, Sato, Ito, Takahashi: "Protease-activated receptor 2 exacerbates adenine-induced renal tubulointerstitial injury in mice." in: Biochemical and biophysical research communications, Vol. 483, Issue 1, pp. 547-552, (2016) (PubMed).

Target Details

Target: Urea Nitrogen (BUN)

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)