QuantiChrom™ Creatinine Assay Kit

Protocol SDS

Application

• For quantitative determination of creatinine and evaluation of drug effects on creatinine metabolism.

Key Features

• Sensitive and accurate. Use 30 µL samples. The detection limit 0.10 mg/dL (8 µM) creatinine in 96-well plate assay.
• Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 5 min. Can be automated as a high-throughput assay for thousands of samples per day.
• Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Assays can be executed in 96-well plate or cuvet.
• Low interference in biological samples. No pretreatments are needed. Assays can be directly performed on raw biological samples.

Method

• OD510nm

Samples

• Urine, serum, plasma and biological preparations

Species

• All

Procedure

• 20 min

Size

• 500 tests

Detection Limit

• 0.1 mg/dL (8 µM)

Shelf Life

• 12 months

More Details

Creatinine is synthesized in the body at a fairly constant rate from creatine, which is produced during muscle contractions from creatine phosphate. In the blood, creatinine is removed by filtration through the glomeruli of the kidney and is secreted into urine. In healthy individuals, creatinine secretion is independent of diet and is fairly constant. The creatinine clearance test has become one of the most sensitive tests for measuring glomerular filtration rate. In kidney disease, creatinine levels in the blood are elevated, whereas the creatinine clearance rate and hence the urine levels are diminished. Creatinine test is most widely used to assess kidney function. Simple, direct and automation-ready procedures for measuring creatinine concentration in biological samples are becoming popular in Research and Drug Discovery. BioAssay Systems creatinine assay kit is designed to measure creatinine directly in biological samples without any pretreatment. The improved Jaffe method utilizes picrate that forms a red colored complex with creatinine. The intensity of the color, measured at 510nm, is directly proportional to creatinine concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw sample.

If I understand correctly, there is no need to perform a full creatinine standard calibration curve. The 2 mg/dL point will suffice. Is this right?

Yes, the reaction is linear from 0 up to 50 mg/dL and a single 2mg/dL creatinine standard is sufficient.

Is this assay compatible with plasma prepared using EDTA?

Yes, EDTA plasma is compatible with the assay.

I have read the specification sheet and I didn’t find details about the temperature of the reaction. Due to it is a kinetic reaction, I am wondering if the 5 minutes incubation should be done at 37°C or room temperature is fine. Do you have some additional information about this?

We normally perform the assay at room temperature but you can run it also at 37°C if this is more convenient. The absorbance values will be higher at 37°C, but because you run the standard and the samples under the same conditions, a change in temperature has no effect on the accuracy of the measurement. It is more important to keep the time between adding the reagent and the OD measurement about the same for all samples, e.g. by using a multichannel pipettor.

I would like to know if the QuantiChrom Creatinine Assay kit can be used for urine samples from rat and mouse.

Yes, this assay can be used for urine samples from any species including rat and mouse.

Ning, L., Suleiman, H. Y., & Miner, J. H. (2020). Synaptopodin is dispensable for normal Podocyte homeostasis but is protective in the context of acute Podocyte injury. Journal of the American Society of Nephrology, 31(12), 2815-2832. Assay: Creatinine in mouse urine.

Casanova, A. G.,et al. (2020). Urinary transferrin pre-emptively identifies the risk of renal damage posed by subclinical tubular alterations. Biomedicine & Pharmacotherapy, 121, 109684. Assay: Creatinine in rat plasma and urine.

Silva Barbosa, A. C.,et al (2020). Inhibition of estrogen Sulfotransferase (SULT1E1/EST) ameliorates ischemic acute kidney injury in mice. Journal of the American Society of Nephrology, 31(7), 1496-1508 Assay: Creatinine in mouse serum.

Kim, J., et al (2020). Protective effects of bee venom against endotoxemia-related acute kidney injury in mice. Biology, 9(7), 154. Assay: Creatinine in mouse plasma.

Mohamed, R.,et al. (2020). Greater high-mobility group box 1 in male compared with female spontaneously hypertensive rats worsens renal ischemia-reperfusion injury. Clinical Science, 134(13), 1751-1762. Assay: Creatinine in rat plasma.

Shen, Y., et al. (2020). Tubule-derived lactate is required for fibroblast activation in acute kidney injury. American Journal of Physiology-Renal Physiology, 318(3), F689-F701. Assay: Creatinine in mouse serum.

Zhang, J.,et al. (2020). Resveratrol decreases high glucose-induced apoptosis in renal tubular cells via suppressing endoplasmic Reticulum stress. Molecular Medicine Reports. Assay: Creatinine in mouse urine.

Cai, T., et al. (2020). Sodium-glucose cotransporter 2 inhibition suppresses HIF-1α-mediated metabolic switch from lipid oxidation to glycolysis in kidney tubule cells of diabetic mice. Cell Death & Disease, 11(5). Assay: Creatinine in human urine.

Weng, W., et al. (2020). Therapeutic effects of fibroblast growth factor-21 on diabetic nephropathy and the possible mechanism in type 1 diabetes mellitus mice. Diabetes & Metabolism Journal, 44(4), 566. Assay: Creatinine in mouse urine.

Tomilin, V. N., et al. (2019). TRPC3 determines osmosensitive [Ca2+]i signaling in the collecting duct and contributes to urinary concentration. PLOS ONE, 14(12), e0226381. Assay: Creatinine in mouse urine.

Kim, J., Leem, J., & Jeon, E. J. (2019). Protective effects of melatonin against Aristolochic acid-induced nephropathy in mice. Biomolecules, 10(1), 11. Assay: Creatinine in mouse plasma.

Kim, J., et al (2020). Inhibition of p300 by Garcinol protects against cisplatin-induced acute kidney injury through suppression of oxidative stress, inflammation, and tubular cell death in mice. Antioxidants, 9(12), 1271. Assay: Creatinine in mouse plasma.

Sun, Z., et al. (2020). Wnt/β-catenin agonist BIO alleviates cisplatin-induced nephrotoxicity without compromising its efficacy of anti-proliferation in ovarian cancer. Life Sciences, 263, 118672. Assay: Creatinine in mouse serum.

Fu, Y., et al (2019). Chronic effects of repeated low-dose cisplatin treatment in mouse kidneys and renal tubular cells. American Journal of Physiology-Renal Physiology, 317(6), F1582-F1592. Assay: Creatinine in mouse serum.

Kim, J., et al. (2020). Protective effects of SPA0355, a thiourea analogue, against lipopolysaccharide-induced acute kidney injury in mice. Antioxidants, 9(7), 585. Assay: Creatinine in mouse plasma.

Svajger, B. A., et al (2020). Development of experimental chronic kidney disease and vascular calcification alters diurnal variation of phosphate and its hormonal regulators. Physiological Reports, 8(21). Assay: Creatinine in rat serum.

Ishak Gabra, et al (2020). Dietary glutamine supplementation suppresses epigenetically-activated oncogenic pathways to inhibit melanoma tumour growth. Nature Communications, 11(1). Assay: Creatinine in mouse serum.

Zhang, J.,et al (2021). Microdose lithium protects against pancreatic islet destruction and renal impairment in streptozotocin-elicited diabetes. Antioxidants, 10(1), 138. Assay: Creatinine in mouse urine.

Zhang, X., et al. (2021). Long non-coding rna rmrp contributes to sepsis-induced acute kidney injury. Yonsei Medical Journal 62(3): 262-273. Assay: Creatinine in mouse serum.

Li, X.-Q., et al. (2019). Deficiency of C3a receptor attenuates the development of diabetic nephropathy. BMJ Open Diabetes Research & Care 7(1): e000817. Assay: Creatinine in mouse urine.

Zhang, J., et al. (2020). Triptolide attenuates renal damage by limiting inflammatory responses in DOCA-salt hypertension. International Immunopharmacology 89(Pt A): 107035. Assay: Creatinine in mouse plasma and urine.

Marek-Bukowiec, K., et al. (2020). The value of urinary RBP4 in the diagnosis of FSGS and other renal diseases. Assay: Creatinine in human urine.

Huang, M., et al. (2019). Integrin-linked kinase deficiency in collecting duct principal cell promotes necroptosis of principal cell and contributes to kidney inflammation and fibrosis. Journal of the American Society of Nephrology: JASN 30(11): 2073-2090. Assay: Creatinine in mouse serum.

Das, S., et al. (2020). Depletion of cyclic-GMP levels and inhibition of cGMP-dependent protein kinase activate p21Cip1 /p27Kip1 pathways and lead to renal fibrosis and dysfunction. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology 34(9): 11925-11943. Assay: Creatinine in mouse urine.

Patel, S., Dhande, I., Gray, E. A., Ali, Q., & Hussain, T. (2019). Prevention of lipopolysaccharide-induced CD11b+ immune cell infiltration in the kidney: role of AT2 receptors. Bioscience reports, 39(5), BSR20190429. Assay: Creatinine in mice plasma/urine.

Jing, W., Jabbari, B., & Vaziri, N. D. (2018). Uremia induces upregulation of cerebral tissue oxidative/inflammatory cascade, down-regulation of nrf2 pathway and disruption of blood brain barrier. American journal of translational research, 10(7), 2137-2147. Assay: Creatinine in rats urine.

Qiu, S., Chen, X., Pang, Y., & Zhang, Z. (2018). Lipocalin-2 protects against renal ischemia/reperfusion injury in mice through autophagy activation mediated by HIF1alpha and NF-kb crosstalk. Biomedicine & Pharmacotherapy, 108, 244-253. Assay: Creatinine in mice serum.

Yin, D. D., Luo, J. H., Zhao, Z. Y., Liao, Y. J., & Li, Y. (2018). Tranilast prevents renal interstitial fibrosis by blocking mast cell infiltration in a rat model of diabetic kidney disease. Molecular medicine reports, 17(5), 7356-7364. Assay: Creatinine in rats kidney cells.

Choi, J. Y., Seth, A., Kashgarian, M., Terrillon, S., Fung, E., Huang, L. & Craft, J. (2017). Disruption of pathogenic cellular networks by IL-21 blockade leads to disease amelioration in murine lupus. The Journal of Immunology, 198(7), 2578-2588. Assay: Creatinine in rats serum.

Chuang, P. Y., Cai, W., Li, X., Fang, L., Xu, J., Yacoub, R. & Lee, K. (2017). Reduction in podocyte SIRT1 accelerates kidney injury in aging mice. American Journal of Physiology-Renal Physiology, 313(3), F621-F628. Assay: Creatinine in mice urine.

Jing, W., Vaziri, N. D., Nunes, A., Suematsu, Y., Farzaneh, T., Khazaeli, M., & Moradi, H. (2017). LCZ696 (Sacubitril/valsartan) ameliorates oxidative stress, inflammation, fibrosis and improves renal function beyond angiotensin receptor blockade in CKD. American journal of translational research, 9(12), 5473-5484. Assay: Creatinine in sprague dewley rats serum.

Liao, H. K., Hatanaka, F., Araoka, T., Reddy, P., Wu, M. Z., Sui, Y. & Guillen, P. (2017). In vivo target gene activation via CRISPR/Cas9-mediated trans-epigenetic modulation. Cell, 171(7), 1495-1507. Assay: Creatinine in mice urine.

West, D., Abou Sawan, S., Mazzulla, M., Williamson, E., & Moore, D. (2017). Whey protein supplementation enhances whole body protein metabolism and performance recovery after resistance exercise: A double-blind crossover study. Nutrients, 9(7), 735. Assay: Creatinine in human urine.

Fujino, T., & Hasebe, N. (2016). Alteration of histone H3K4 methylation in glomerular podocytes associated with proteinuria in patients with membranous nephropathy. BMC nephrology, 17(1), 179. Assay: Creatinine in mouse cells.

Min, D., Brooks, B., Wong, J., Aamidor, S., Seehoo, R., Sutanto, S. & McLennan, S. V. (2016). Monocyte CD163 is altered in association with diabetic complications: possible protective role. Journal of leukocyte biology, 100(6), 1375-1383. Assay: Creatinine in mice plasma.

Pai, C. H., Yen, C. T., Chen, C. P., Yu, I. S., Lin, S. W., & Lin, S. R. (2016). Lack of Thromboxane synthase prevents hypertension and fetal growth restriction after high salt treatment during pregnancy. PloS one, 11(3), e0151617. Assay: Creatinine in mice urine.

Zheng, S., Coventry, S., Cai, L., Powell, D. W., Jala, V. R., Haribabu, B., & Epstein, P. N. (2016). Renal protection by genetic deletion of the atypical chemokine receptor ACKR2 in diabetic OVE mice. Journal of diabetes research, 2016:5362506. Assay: Creatinine in mice serum.

Zhou, X., Chen, K., Wang, Y., Schuman, M., Lei, H., & Sun, Z. (2016). Antiaging gene Klotho regulates adrenal CYP11B2 expression and aldosterone synthesis. Journal of the American Society of Nephrology, 27(6), 1765-1776. Assay: Creatinine in mice urine.

Hauser PV, et al (2010). Stem cells derived from human amniotic fluid contribute to acute kidney injury recovery. Am J Pathol. 177(4):2011-21. Assay: Creatinine in mice serum.

Jaruga, P. and Dizdaroglu, M (2010). Identification and quantification of (5’R)- and (5’S)-8,5′-cyclo-2′-deoxyadenosines in human urine as putative biomarkers of oxidatively induced damage to DNA. BBRC 397(1): 48-52. Assay: Creatinine in human urine.

Kim JW, et al (2010). Increased urinary lipocalin-2 reflects matrix metalloproteinase-9 activity in chronic hepatitis C with hepatic fibrosis. Tohoku J Exp Med. 222(4):319-27. Assay: Creatinine in human urine.

Sahan-Firat, S et al (2010). 2,3′,4,5′-Tetramethoxystilbene prevents deoxycorticosterone-salt-induced hypertension: contribution of cytochrome P-450 1B1. Am J Physiol Heart Circ Physiol 299(6):H1891-901. Assay: Creatinine in rat serum.

Tang TC,et al (2010). Impact of metronomic UFT/cyclophosphamide chemotherapy and antiangiogenic drug assessed in a new preclinical model of locally advanced orthotopic hepatocellular carcinoma. Neoplasia 12(3):264-74. Assay: Creatinine in mice urine.

Xu J, et al (2010). FVB mouse genotype confers susceptibility to OVE26 diabetic albuminuria. Am J Physiol Renal Physiol. 299(3):F487-94. Assay: Creatinine in mouse serum.

Andonegui G, et al (2009). Characterization of S. pneumoniae pneumonia-induced multiple organ dysfunction syndrome: an experimental mouse model of gram-positive sepsis. Shock. 31(4):423-8. Assay: Creatinine in mouse plasma.

Zafar I, et al (2009). Long-term rapamycin therapy in the Han:SPRD rat model of polycystic kidney disease (PKD). Nephrol Dial Transplant. 24(8):2349-53. Assay: Creatinine in rat serum.

Gilliam-Davis S, et al (2007). Long-term AT1 receptor blockade improves metabolic function and provides renoprotection in Fischer-344 rats. Am J Physiol Heart Circ Physiol. 293(3):H1327-33. Assay: Creatinine in rat serum.

Lorch, G et al (2007). Inhibition of epidermal growth factor receptor signalling reduces hypercalcaemia induced by human lung squamous-cell carcinoma in athymic mice. Br J Cancer 97(2):183-93. Assay: Creatinine in mouse urine.

Stanic, AK et al (2006). Immune dysregulation accelerates atherosclerosis and modulates plaque composition in systemic lupus erythematosus. PNAS 103(18):7018-23. Assay: Creatinine in mouse serum.

Zhang SX, et al (2006). Therapeutic potential of angiostatin in diabetic nephropathy. J Am Soc Nephrol. 17(2):475-86. Assay: Creatinine in rat urine.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.