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Total Collagen Assay Kit (Colorimetric)

A simple, sensitive & easy colorimetric assay to quantitate total collagen levels.
Catalog #: K218

Product Details

Cat # +Size K218-100
Size 100 assays
Detection Method Absorbance (OD = 560 nm).
Species Reactivity Mammalian tissues, protein/peptide hydrolysates, serum, urine
Applications Measurement of collagen in various sample types.
Features & Benefits • Simple, fast and convenient assay
• Can detect as low as 0.5 µg of collagen in a variety of samples
Kit Components • Oxidation Buffer
• Chloramine T Concentrate
• Perchloric Acid/Isopropanol Solution
• DMAB Concentrate (in DMSO)
• Collagen I Standard (2 mg/ml)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Collagen is the most abundant insoluble protein found in the extracellular matrix and connective tissues. It can be found in skin, tendons, bone, cartilage, muscle, vitreous humor and ligaments, among other tissues. There are more than sixteen - well characterized types of collagens, but types I, II and III collagen comprise more than 80% content in mammals. The triple-helical structure of collagen is quite unique: it consists of a repeating pattern of a basic trimer: Glycine-Proline-Hydroxyproline. In cells, collagens are secreted as procollagens and these chains are transported into the Endoplasmic Reticulum, where, numerous post-translational modifications lead to the formation of a triple helix with disulfide bonds. Excessive production of collagen is linked to pathological conditions including liver cirrhosis, lung fibrosis, and tumor growth. BioVision’s Collagen Assay Kit is a simple and sensitive assay to detect small amounts of collagens in a variety of samples. The assay is based on the acid hydrolysis of samples to form hydrolysates and Hydroxyproline. This released Hydroxyproline gets oxidized to form a reaction intermediate, which further in the reaction, forms a chromophore (Abs 560 nm). The assay is simple, sensitive and specific for collagen and can detect as low as 0.5 µg of collagen in a variety of samples such as tissue homogenates, biological fluids and purified proteins.

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What is a suitable positive control for this assay?
The standard can be used as a positive control to ensure that the kit components are working well.
Could you please suggest our customer the method of serum sample preparation?
A typical protocol for serum preparation is as follows:
1. Collect blood using any anticoagulant
2. Allow the blood to clot for 30 min at 25C
3. Centrifuge at 2000g for 15min at 25C. Pipette off the top yellow serum layer without disturbing the white buffy layer
4. Proceed with 100ul of the sample
What is the source of collagen in the kit?
The collagen included in the kit is isolated from rat tail.
Could you let us know why collage type I (K218-100-5) is used as standards in this kit?
Collagen type I is the most abundant form of collagen found in human tissues and that’s why it has been incorporated as a standard in the kit.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Will the kit measure pro-collagen?
The kit only measures Collagen. All the Collagen in your sample is hydrolyzed to hydrolysates and hydroxyproline. Hydroxyproline is then oxidized to an intermediate which forms a chromophore detected at 560 nm.
Does glutaraldehyde crosslinking of the collagen fibers change the assay outcome?
Glutaraldehyde cross linking may not be stable enough and acid hydrolysis should be enough to break down the collagen theoretically. However, we have not tested this.
We have a lot of samples and we are not able to process them all at the same time. I was wondering at what stage in sample processing can I stop and freeze them? Can we freeze our samples after we hydrolyze them?
Yes, you can definitely freeze your samples at -20°C or -80°C after hydrolysis. Hydroxyproline is fairly stable and should be good for a week to 10 days at -80°C. We do not recommend long term storage of hydrolyzed samples as freshly frozen samples are always better than samples frozen for months.
Ute Nazet. Mechanical Stress Induce PG-E2 in Murine Synovial Fibroblasts Originating from the Temporomandibular Joint. Cells. Feb 2021. 33535605.
Jongmin Kim. Development of 3D Printed Bruch’s Membrane-Mimetic Substance for the Maturation of Retinal Pigment Epithelial Cells. Int J Mol Sci. Jan 2021. 33499245.
For more citations of this product click here