Thank you!

Your quote has been successfully submitted!

For products requiring additional information, our team will contact you within 1 business day

Failed

There was an error submitting your quote. Please try again.

Hdroxyproline (HYP) Colorimetric Assay Kit– MSE Supplies LLC

Overstock Sale - Select Products 10% Off on Orders of $500 or More! Promo Code:

SAVING10

Shop Now
Menu

Hdroxyproline (HYP) Colorimetric Assay Kit

SKU: E-BC-K062-S-100

To better serve you, we would like to discuss your specific requirement.
Please Contact Us for a quote.

Hdroxyproline (HYP) Colorimetric Assay Kit

SKU # E-BC-K062-S
Detection Instrument Spectrophotometer (550-570 nm, optimum wavelength: 558 nm)
Detection Method Colorimetric method


Product Details

Properties

Synonyms HYP
Sample Type Serum, animal tissue and urine samples
Sensitivity 0.032 μg/mL
Detection Range 0.032-10 μg/mL
Detection Method Colorimetric method
Assay type Quantitative
Assay time 45 min
Precision Average inter-assay CV: 5.700% | Average intra-assay CV: 4.900%
Other instruments required Vortex mixer, Centrifuge, Water bath
Other reagents required 6 mol/L Hydrochloric acid, Concentrated hydrochloric acid (12 mol/L), N-propyl alcohol
Storage 2-8℃
Valid period 12 months


Dilution of Sample

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

Sample type Dilution factor
10% Rat liver tissue homogenate 1
10% Rat kidney tissue homogenate 1
10% Rat lung tissue homogenate 1
10% Rat brain tissue homogenate 1
Chicken Tendon 20-30
Fish scale 20-30
Porcine cartilage 15-25
Chicken cartilage 15-25
Human Urine 1
Fetal bovine serum 1
Rat plasma 1

 

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

Item Component Size 1 (50 Assays) Size 2 (100 Assays) Storage
Reagent 1 Oxidant Agent Powder × 1 vial Powder × 1 vial 2-8℃, 12 months,
shading light
Reagent 2 Buffer Solution 8 mL × 1 vial 15 mL × 1 vial 2-8℃, 12 months
Reagent 3 Oxidant Agent
Solvent
8 mL × 1 vial 15 mL × 1 vial 2-8℃, 12 months
Reagent 4 Chromogenic Agent Powder × 1 vial Powder × 1 vial 2-8℃, 12 months,
shading light
Reagent 5 Chromogenic Agent Solvent 28 mL ×1 vial 52 mL ×1 vial 2-8℃, 12 months
Reagent 6 Standard 5 mg × 1 vial 5 mg × 2 vials 2-8℃, 12 months,
shading light
Reagent 7 pH Adjusting Solution A 60 mL × 1 vial 60 mL × 2 vials 2-8℃, 12 months
Reagent 8 pH Adjusting Solution B 60 mL × 1 vial 60 mL × 2 vials 2-8℃, 12 months
Reagent 9 Clarificant Powder × 1 vial Powder × 2 vials 2-8℃, 12 months


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).

Parameters Sample 1 Sample 2 Sample 3
Mean (μg/mL) 0.80 2.60 6.80
%CV 5.3 4.8 4.6


Inter-assay Precision

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

Parameters Sample 1 Sample 2 Sample 3
Mean (μg/mL) 0.80 2.60 6.80
%CV 5.6 6.0 5.5


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%.


Standard 1 Standard 2 Standard 3
Expected Conc. (μg/mL) 2.5 5.7 8.3
Observed Conc. (μg/mL) 2.7 5.8 8.8
Recovery rate (%) 106 102.4 106.3


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:

Concentration (μg/mL) 0 2 3 4 6 8 10
OD Value 0.005 0.140 0.210 0.278 0.413 0.536 0.669
0.004 0.146 0.204 0.294 0.410 0.496 0.645
Average OD 0.005 0.143 0.207 0.273 0.412 0.516 0.657
Absoluted OD 0.000 0.139 0.203 0.269 0.407 0.512 0.653