Type
Colorimetric assay
Description
Now collagen can be easily determined in small tissue samples with low collagen levels!
The Sensitive Tissue Collagen assay kit is based on the quantitative colorimetric determination of hydroxyproline residues, obtained by acid hydrolysis of collagen. The Sensitive Tissue Collagen assay shows no matrix effect and no need for further dilution of the hydrolysate enabling measuring in small samples with low collagen concentrations, such as liver. The assay is simple and it doesn’t need the drying step following acid hydrolysis, resulting in a fast (<2 hrs) and easy (96-well plate format) assay following the hydrolysis step.
Collagen is one of the main components of extracellular matrix. Dysregulation in collagen production results 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.
The QuickZyme Sensitive Tissue Collagen assay is based on the detection of hydroxyproline. Hydroxyproline is a non-proteinogenic amino acid, which in mammals occurs mainly in collagen. Hydroxyproline in tissue hydrolysates can be used as a direct measure of the amount of collagen present in the tissue.
The measurement of collagen is started by complete hydrolysis of tissue samples in 6M HCl at 95°C. In the hydrolysate hydroxyproline residues are quantified using 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, which 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.
In contrast to other hydroxyproline-based assays, the QuickZyme Sensitive Tissue Collagen assay has no matrix effect and no need for dilution of the hydrolysate enabling measuring in small samples with low collagen concentrations. The assay is simple and it doesn’t need the drying step following acid hydrolysis for which often special equipment is needed.
Applications
Tissue, Tissue homogenates
Sample Requirements
50 - 500 µl
Shipping
At ambient temperature. Upon receipt, store the product at the temperature recommended below.
Storage/Expiration
Unopened kit: Store at room temperature (RT) in the dark. Do not use kit components past kit expiration date.
Opened kit: reconstituted reagents: The opened collagen 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.
Calibration Range
0 - 300 µg/ml
Limit of Detection
2.5 µg/ml