Type
Colorimetric assay
Description
Now hydroxyproline can be easily determined in small tissue samples with low collagen levels!
Dysregulation in collagen metabolism may result in pathologies such as fibrosis (too much collagen), or osteoarthritis (too little collagen). Therefore measurement of collagen production is important in many disease related studies.
Hydroxyproline is a non-proteinogenic 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 QuickZyme Sensitive Tissue Hydroxyproline assay is a modification of the method described by Prockop and Udenfriend (Anal. Biochem.,1960, 1: 228-239). 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 QuickZyme Sensitive Tissue Hydroxyproline assay kit is the only hydroxyproline-based assay which shows no matrix effect and no need for further dilution of the hydrolysate enabling measurement in small samples with low hydroxyproline concentrations. In addition, no drying of hydrolysate is needed, resulting in a fast (<2 hrs), sensitive and easy (96-well plate format) assay.
Applications
Tissue, Tissue homogenates
Sample Requirements
5 - 35 μl
Shipping
At ambient temperature. Upon receipt, store the product at the temperature recommended below.
Storage/Expiration
Unopened kit: Store at room temperature in the dark. Do not use kit components past the kit expiration date.
Opened kit / reconstituted reagents: The opened standard, assay buffer and enhancer solution should be stored light protected at 4°C. The other opened reagents should be stored light protected at RT and are stable for at least 1 month. The reconstituted detection reagent (A+B) should be used on the day of reconstitution.
Calibration Range
0 - 300 µM
Limit of Detection
2.4 µM (0.3 µg/ml)
Note
NOTE: 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