Creatinine Microplate Assay Kit


The Creatinine Microplate Assay kit is for the quantitative determination of Creatinine in urine by a microplate assay. The Creatinine Microplate Assay Kit is for research use only and not for use in diagnostic procedures.

SKU: CRE34-K01 Categories: , ,

Creatinine Microplate Assay Kit

The Creatinine Microplate Assay Kit is For Research Use Only

Size: 1×96 wells
Sensitivity: 1.0 mg/dL
Dynamic Range: 1.0 – 10 mg/dL
Incubation Time: 15 minutes
Sample Type: Urine
Sample Size: 25 µl
Alternative Names: Cr, Creatine
For Research Use Only

Assay Background for Creatinine Microplate Assay

Creatine (Cr) is produced in the kidney, liver and pancreas, phosphorylated, and transported to the brain and muscle tissue. However, a small proportion of free Cr is converted irreversibly to creatinine (Crn) in the muscular tissue in proportion to the muscle mass. The amount of Crn excreted daily by an individual is relatively constant.  Thus, urinary creatinine levels may be used as an index of standardization. 24-h urinary Crn excretion is used to estimate total muscle mass because the rate of non-enzymatic production of Crn from Cr is nearly constant and >90% of the total body Cr is found in muscle tissue. Normal urinary creatinine values for men and women range from 9.7 – 24.7 and 7.9 – 14.2 mmol/24h respectively. Changes in excretion rate may be indicative of impaired renal metabolism.

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Additional Information

Assay Principle

The Creatinine Microplate Assay is a colorimetric assay for the quantitative analysis of creatinine levels in urine. Urinary creatinine reacts with picric acid under alkaline conditions to produce an orange color, which can be quantified by absorption spectroscopy near the 500 nm wavelength. This reaction, known as the Jaffe reaction, also occurs non-specifically with other components in biological fluids. However, the specific color produced with creatine in this reaction is known to degrade rapidly under acidic conditions (Slot et al.). Heinegard and Tiderstrom showed that the difference in color intensity determined before and after the addition of acid is a direct estimate of creatinine concentration in the sample.

  1. Add 25 µL of Standards or Samples (may require diluting 1:4 or 1:8) to the corresponding wells on the microplate in duplicate.  See Scheme I for a sample plate layout.
  2. Add 180 µL of the Alkaline Picrate Reagent to each well.
  3. Mix by shaking or placing the plate on a shaker and incubate at room temperature for 10 minutes.
  4. Read the plate at 490 nm. (First Reading)
  5. Add 15 µL of R3: Acid Reagent to each well.
  6. Mix thoroughly by tapping or shaking and allow to stand at room temperature for 5 minutes.
  7. Read the plate again at 490 nm. (Second Reading)

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