EnzyChrom™ Creatinine Assay Kit
Protocol SDS

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

  • All

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 λex/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 standard
2) 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, 5 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 OD570nm.

The sample creatinine concentration is computed as follows:
[Creatinine] = (RSAMPLE – RBLANK)/(RSTANDARD – RSAMPLE) x [Standard]/4 x n (µM)

where RSAMPLE, RBLANK, and RSTANDARD 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.

- Fast turnaround
- Quality data
- Low cost

Please email or call 1-510-782-9988 x 2 to request assay service.
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EnzyChrom™ Creatinine Assay Kit
Catalog No: EICT-100
Price: $329    Qty:
For orders of 10 or more kits, please call +1-510-782-9988x1 or email us for best pricing and/or bulk order.

Shipping: On Ice
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: -20°C

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