Hdroxyproline (HYP) Colorimetric Assay Kit

Hdroxyproline (HYP) Colorimetric Assay Kit

Product Details

Properties

Dilution of Sample

The recommended dilution factor for different samples is as follows (for reference only):

Note: The diluent is double distilled water (For little tissue sample, the addition of hydrochloric acid solution, pH adjustment solution and final constant volume can be reduced proportionally. At least 400 μL of sample hydrolysate is required for detection). For the dilution of other sample types, please do pretest to confirm the dilution factor.

Detection Principle

The sample is hydrolyzed to generate free HYP, and hydroxyproline can produce oxidation product under the action of oxidizing agent. The generated oxidation product can react with chromogenic agent to produce burgundy. The concentration of hydroxyproline can be calculated by measuring the OD value at 558 nm.

Kit Components & Storage

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 104.9%.

Sensitivity

The analytical sensitivity of the assay is 0.032 μg/mL. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.

Standard Curve:

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only: