This kit is for the quantitative measurement of hydroxyproline and is optimized for tissues with low collagen or hydroxyproline content (such as liver) due to absence of matrix effects. This assay has the same application area as the total collagen assay, but is intended for customers who have their own method of acid hydrolysis, or have a collection of hydrolyzed samples to be analyzed.
Also available in 5x 96-assays bulk pack (QZBtishyp5)
Hydroxyproline is a nonproteinogenic amino acid, which in mammals occurs in elastin and collagen. Its presence is mainly limited to the triple helix of collagen, where it leads to increased triple helix stability. Hydroxyproline is formed post translationally from specific proline residues by action of the enzyme prolylhydroxylase. Hydroxyproline in tissue hydrolysates can be used as a direct measure of the amount of collagen present.
The assay measures the total amount of hydroxyproline present in the sample. If this hydroxyproline is obtained upon hydrolysis of collagen, it represents all the types of collagen present in the sample without discriminating between the types of collagen and between procollagen, mature collagen and collagen degradation products. The assay is simple and results in a chromogen with a maximum at 570 nm. The assay is developed absorbance to measure hydroxyproline in acid tissue and/or protein hydrolysates in such a way that it doesn’t need the drying step following acid hydrolysis collagen for which often special equipment is needed.
This kit does not contain tubes and a protocol for collagen hydrolysis. If you need these items please order the QuickZyme sensitive tissue collagen assay.
Each 96 well kit cotains:
- 2x adhesive plate seals
- Assay buffer
- 2x Detection reagent A
- Detection reagent B
- Hydroxyproline standard (3 mM) in sterile water
- 2x Enhancer solution
- 96-well assay plate
- Assay protocol booklet
The following materials and equipment are required but not supplied:
- 4M HCl for sample and standard dilution
- MilliQ or comparable high quality water
- Single and/or multichannel pipettes
- Eppendorf centrifuge
- Incubator for heating at 60°C
- Microplate reader capable of measuring at a wavelength between 540 and 580 nm, 570 nm preferred.
- Microplate shaker
- PP-, PE- or glass tubes (no polystyrene)
- Eppendorf tubes
This kit does not contain tubes and a protocol for collagen hydrolysis.
If you need these items please order the QuickZyme sensitive tissue collagen assay.
Samples: tissue homogenates, tissues
Range: 6 to 300 μM.
Sensitivity: 2.4 µM (0.3 µg/ml)
Uses hydroxyproline as standard
Ease-of-use: Equivalent to ELISA
Colorimetric read out
Run time: < 2 hours
Storage at room temperature
Application note 1: How to choose your collagen assay