EnzyChrom™ Creatinine Assay Kit

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ProtocolSDS

Application

Creatinine determination in urine, serum, plasma, and other biological preparations.

Key Features

Fast and sensitive. Linear detection range: 10.6 to 500 µM or 0.12-5.7 mg/dL (colorimetric assay) and 0.25 to 50 µM or 0.0028-0.57 mg/dL (fluorimetric assay) for a 60 min reaction.
Convenient. The procedure involves adding a single working reagent and reading after 60 minutes.
High-throughput. Homogeneous "mix-incubate-measure" type assay. Can be readily automated to process thousands of samples per day.

Method

OD570nm; FL530/585nm

Samples

Urine, serum, plasma, and other biological preparations.

Species

Size

100 tests

Detection Limit

Colorimetric assay: 10.6 µM or 0.12 mg/dL; Fluorimetric assay: 0.25 µM or 0.0028 mg/dL

Shelf Life

6 months

More Details

CREATININE is synthesized in the body 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 testing is most widely used to assess kidney function.BioAssay Systems’ enzyme-based creatinine assay uses a reaction sequence that excludes both endogenous creatine and ammonia to convert a dye into a colored and fluorescent form. The absorbance at 570 nm or fluorescence intensity at λ_ex/em = 530/585 nm is directly proportional to the creatinine concentration in the sample.

What samples have you tested?

This kit has been tested and validated in urine, plasma, and serum samples.

Can I store unused reagents for future use?

Yes, unused reagents can be stored according to the assay protocol.

How do you use this kit on deproteinated plasma or serum samples with an internal standard procedure?

Samples will need three separate reactions: 1) sample plus standard2) sample alone 3) sample blank. For the internal standard prepare 200 µL 1250 µM standard by mixing 125 µL 2 mM Standard and 75 µL dH2O. For the sample plus standard well, add 5 µL 1250 µM standard and 20 µL sample. For the sample and sample blank wells, add 5 µL dH2O and 20 µL sample. Prepare enough Working Reagent (WR) for all assay wells by mixing, for each well, 80 µL Assay Buffer, 1 µL Enzyme A, 1 µL Enzyme Mix, 1 µL ATP, 1 µL Dye Reagent. Prepare a WR without the Enzyme A for the sample blank well(s). Add 80 µL of the WR to each well. Tap plate briefly to mix. Incubate at room temperature for 60 min. Use a plate reader to read OD_570nm. The sample creatinine concentration is computed as follows: [Creatinine] = (R_SAMPLE – R_BLANK)/(R_STANDARD – R_SAMPLE) x [Standard]/4 x n(µM)where R_SAMPLE, R_BLANK, and R_STANDARD are the OD or fluorescence values of the Sample, Sample Blank, and the Sample plus Standard respectively. n is the sample dilution factor. Note: The volume of the internal standard is 4× lower than the sample volume; thus, the internal standard concentration should be divided by 4.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.

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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.Simply send us your samples:- Fast turnaround - Quality data - Low costPlease email or call 1-510-782-9988 x 2 to request assay service.

品牌介绍

BioAssay Systems开发和销售创新的高通量分析解决方案，以满足生命科学行业不断增长的需求。BioAssay Systems的创始人在现代药物发现领域拥有50多年的丰富经验。在过去的15年中，我们已经开发了200多种用于药物发现和生命科学研究的检测试剂盒。我们以提供的服务和简单，易用且性能卓越的产品而感到自豪。

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