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QuickZyme Total Collagen Assay Kit

  • Regulatory status:RUO
  • Type:Colorimetric assay
  • Species:Multispecies
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Cat. No. Size Price


New QZBTOTCOL1 96 wells (1 kit)
New QZBTOTCOL2 2 x 96 wells (1 kit)
New QZBTOTCOL5 5 x 96 wells (1 kit)
PubMed Product Details
Technical Data

Type

Colorimetric assay

Description

Now collagen can be easily determined by any biochemical laboratory!

The QuickZyme Total Collagen assay kit is based on the quantitative colorimetric determination of hydroxyproline residues, obtained by acid hydrolysis of collagen. Usually for this analysis upon hydrolysis removal of HCl is needed, which is a time-consuming step and/or needs special equipment. The QuickZyme Total Collagen assay is the only hydroxyproline-based assay which does not need this laborious step, 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 Total Collagen assay is based on the detection of hydroxyproline. 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 its presence increases the triple helix stability. Hydroxyproline is formed posttranslationally 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 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.

The assay is simple and results in a chromogen with an absorbance maximum at 570 nm. The assay is developed in such a way that it doesn’t need the drying step following acid hydrolysis for which often special equipment is needed.

Applications

Tissue, Cellular extracts, 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

Summary

Features

  • Quantitative measurement of all types of collagen, species independent.

  • Samples: Cellular extracts, tissue homogenates, tissues.

  • Colorimetric measurement of hydroxyproline content.

  • Range: 6 to 300 μg/ml. Sensitivity: 2.5 µg/ml.

  • Uses rat tail collagen as standard .

  • Ease-of-use: Equivalent to ELISA .

  • Store at room temperature

Research topic

Extracellular matrix

Summary

Collagens are the most abundant proteins in mammals. The collagen family comprises 28 members that contain at least one triple-helical domain. Collagens are deposited in the extracellular matrix where most of them form supramolecular assemblies. Four collagens are type II membrane proteins that also exist in a soluble form released from the cell surface by shedding. Collagens play structural roles and contribute to mechanical properties, organization, and shape of tissues. They interact with cells via several receptor families and regulate their proliferation, migration, and differentiation. Some collagens have a restricted tissue distribution and hence specific biological functions.

Summary References (1)

References to Collagen

  • Ricard-Blum S. The collagen family. Cold Spring Harb Perspect Biol. 2011 Jan 1;3(1):a004978. doi: 10.1101/cshperspect.a004978. PMID: 21421911; PMCID: PMC3003457. See more on PubMed
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