Quantitative hydroxyproline measurement for collagen analysis in all types of tissues, in a fast & simple 96-well plate format!
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 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 assay is simple and results in a chromogen with an absorbance maximum at 570 nm. The assay is developed 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 of collagen for which often special equipment is needed. The QuickZyme hydroxyproline assay kit is the only hydroxyproline-based assay which does not need this laborious step, resulting in a fast (<2 hrs), sensitive and easy (96-well plate format) assay.
Tissue, Cellular extracts, Tissue homogenates, Acid hydrolized tissue culture supernatants
5 - 35 µl
At ambient temperature. Upon receipt, store the product at the temperature recommended below.
Unopened kit: Store at room temperature in the dark. Do not use kit components past kit expiration date.
Opened kit / reconstituted reagents: The opened standard and assay buffer 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.
0 - 300 µM
Limit of Detection
2.4 µM (0.3 µg/ml)
This kit does not contain tubes or a protocol for collagen hydrolysis. If you need these items please order the QuickZyme Total Collagen Assay