Specificity:
The RayBio Creatinine assay is designed to measure Creatinine in urine.
Number of Targets Detected:
1
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Solid Support:
96-well Microplate
Size:
2 or 4 x 96-Well Strip Microplate Kit
Creatinine is a metabolite waste product of phospho-Creatine (p-Creatine), a molecule used as a store for high-energy phosphate that can be utilized by muscles and other tissues for the production of ATP. Dietary intake andde novo synthesis from the amino acids arginine, glycine, and methioninein the kidney and liver are the primary sources of Creatine. Creatine and p-Creatine are converted non-enzymatically to the metabolite Creatinine, which diffuses into the blood, is filtered through the kidneys, and is then secreted into urine. In vivo, this conversion appears to be irreversible and in vitro it is favored by higher temperatures and lower pH. Some Creatinine may form via p-Creatinine as well. Under normal conditions, Creatinine's formation occurs at a rate that is relatively constant. For instance, in humans approximately 2% of the Creatine/p-Creatine pool is converted to Creatinine daily. This predictability makes Creatinine a useful tool for normalizing the levels of other molecules found in urine. In addition, altered Creatinine levels can be used as an indicator of kidney dysfunction, or may be associated with other conditions that result in decreased renal blood flow. Some examples include diabetes and cardiovascular disease. The RayBio® Creatinine assay is a 30 minute chemical analysis designed to measure Creatinine in urine.
Kit Components
- 96-well Strip Microplate
- Creatinine Standard
- Picric Acid Reagent
- NaOH
Other Materials Required
- Microplate reader capable of measuring absorbance at 490 nm
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Distilled or deionized water
- Tubes to prepare standard or sample dilutions
Protocol Outline
- Bring all reagents and samples to room temperature before use. It is recommended that all standards and samples be run at least in duplicate.
- Add 50 µL standard and sample to appropriate wells.
- Add 100 µL Alkaline Picrate Solution to each well. Incubate for 30 minutes at room temperature.
- Read at 490 nm immediately.
The entire kit should be stored at 2-8 °C for up to 6 months from the date of shipment. Do not use past kit expiration date.