Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Tissue Homogenate, Serum,
Assay type:Quantitative,
Range:0.5 - 10 µg,
Assay time:3h,
Assay Platform:Microplate reader
Product details:
Total collagen assay principle
The total collagen assay protocol is based on the total alkaline/acid hydrolysis of samples to breakdown the collagen trihelix peptide into single amino acids and yield free Hydroxyproline. The released hydroxyproline is oxidized to form a reaction intermediate, which further in reaction forms brightly-colored chromophore that can be easily detected at OD 560 nm.
The total collagen assay uses an identical assay principle and assay components to our hydroxyproline assay kit
ab222941
, except that the standard curve of this assay is constructed from collagen itself (and that collagen is treated in an identical manner to samples, undergoing alkaline hydrolysis at 120C). This makes this assay a more direct way to quantify collagen levels.
The assay employs a proprietary developer solution to accurately measure collagen in hydrolysates without the use of hazardous perchloric acid, removing the need for any special handling and waste-disposal protocols.
Total Collagen assay kit protocol summary:
- Hydrolyze samples and standard with concentrated NaOH for 1 hr at 120°C
- Cool on ice and neutralize with concentrated HCL
- Centrifuge at 10,000g for 5 min to pellet debris
- Add samples and standards to wells
- Evaporate wells to dryness at 65°C
- Add oxidation mix and incubate for 20 min
- Add developer and incubate for 5 min
- Add DMAB concentrate and incubate for 45 min at 65°C
- Analyze absorbance at 560 nm with a microplate reader
How other researchers are using
Total Collagen Assay Kit (Perchlorate-Free) has been used in a variety of sample type including:
- Mouse liver samples
- Mesenchymal stem cell culture
References:
1- Chen X al. 2023
2 Gousopoulou X et al. 2023
This product is manufactured by BioVision, an Abcam company and was previously called K406 Total Collagen Assay Kit (Perchlorate-Free). K406-100 is the same size as the 100 test size of ab222942.
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 Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information-Please refer to protocols
