Hydroxyproline Assay Kit Colorimetric ab222941 provides a quick and convenient method to quantify hydroxyproline in tissue lysates and biological fluids such as urine and serum.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Publications
- Chinese medicine 19:102024Modified Qing-Zao-Jiu-Fei decoction attenuated pulmonary fibrosis induced by bleomycin in rats via modulating Nrf2/NF-κB and MAPKs pathways.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJia-Qian Zhu,Yuan-Yang Tian,Kam Leung Chan,Zhen Hu,Qing-Qing Xu,Zhi-Xiu Lin,Yan-Fang XianPubMed 38229198
- Scientific reports 13:205772023Superiority of systemic bleomycin to intradermal HOCl for the study of interstitial lung disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesArina Morozan,Sydney Joy,Utako Fujii,Richard Fraser,Kevin Watters,James G Martin,Inés ColmegnaPubMed 37996447
Key facts
- Detection method
Colorimetric
- Sample types
Urine, Tissue Homogenate, Serum, Other biological fluids
- Assay type
Quantitative
- Reactive species
Mammals
- Range
0.1 - 1 µg
- Assay time
3h
Target data
What's included?
Recommended products
Hydroxyproline Assay Kit Colorimetric ab222941 provides a quick and convenient method to quantify hydroxyproline in tissue lysates and biological fluids such as urine and serum.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Key facts
- Detection method
Colorimetric
- Sample types
Urine, Tissue Homogenate, Serum, Other biological fluids
- Assay type
Quantitative
- Reactive species
Mammals
- Range
0.1 - 1 µg
- Assay time
3h
- Assay Platform
Microplate reader
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
Multi
- Storage information
Please refer to protocols
Notes
Hydroxyproline Assay Kit (Colorimetric) ab222941 provides a quick and convenient method to quantify hydroxyproline in tissue lysates and biological fluids such as urine and serum.
The classical hydroxyproline assay protocol is based on the oxidation of hydroxyproline to a pyrrole intermediate followed by reaction with Ehrlich's reagent dissolved in concentrated perchloric acid. Perchloric acid is a hazardous material that is both toxic and highly reactive, requiring special handling and waste-disposal protocols.
This hydroxyproline assay protocol employs a proprietary acidic developer solution to accurately measure hydroxyproline in hydrolysates without the use of hazardous perchlorates. It is a quick and convenient protocol where hydroxyproline gets oxidized to form a reaction intermediate, which further in reaction forms brightly-colored chromophore that can be easily detected at OD 560 nm.
The assay can detect as low as 0.05 μg hydroxyproline/well.
This product is a replacement for Hydroxyproline Assay Kit K555; this kit does not contain the hazardous Perchloric acid which is present in K555.
The components in this product are exactly the same as in K555, except that the Perchloric acid/Isopropanol Solution component in K555 has been replaced with the Developer component.
If you would prefer to continue to use a kit in the format of K555, then use this product and:
For safety reasons, Perchloric Acid will not be shipped by Abcam in the same shipment as either Isopropanol or this product.
1) In a fume cupboard, mix 1 ml of 70% Perchloric Acid and 5 ml of Isopropanol to produce the Perchloric acid/Isopropanol Solution component previously provided in K555.
2) Then follow the protocol for K555 available on www.biovision.com/documentation/datasheets/K555.pdf, using the components from this product, and the Perchloric acid/Isopropanol Solution that you produced.
This product is manufactured by BioVision, an Abcam company and was previously called K226 Hydroxyproline Assay Kit (Perchlorate-Free). K226-100 is the same size as the 100 test size of ab222941.
Hydroxyproline assay protocol summary:
- add 10 N concentrated NaOH to samples and hydrolyze at 120°C for 1 hr
- cool on ice
- neutralize with 10 N concentrated HCl, centrifuge and collect supernatant
- add samples and standards to wells
- evaporate wells to dryness by heating at 65°C
- add oxidation reagent mix to dissolve crystalline residue, and incubate at room temp for 20 min
- add developer and incubate at 37°C for 5 min
- add DMAB concentrate and incubate for 45 min at 65°C
- analyze with microplate reader
We are currently not able to offer K555 Hydroxyproline Colorimetric Assay Kit from BioVision and apologize for the inconvenience that this may cause. We recommend ab222941 as an alternative.
Supplementary info
- Activity summary
Hydroxyproline also known as Hyp is a non-standard amino acid with a molecular weight of approximately 131.13 g/mol. It is primarily found in collagen making up about 13% of its structure. Hydroxyproline expression is mainly localized in tissues where collagen is abundant such as skin bone and cartilage. This makes it an important marker in studies related to connective tissue health and integrity and therefore hydroxyproline assays and kits are commonly used for analyzing collagen content in these tissues.
- Biological function summary
Hydroxyproline plays a critical role in stabilizing the triple helical structure of collagen. It is not directly encoded by the genetic code but formed by the post-translational modification of proline residues in collagen chains. The presence of hydroxyproline in collagen is essential as it ensures the structural stability and mechanical strength of the collagen thereby supporting tissue integrity. This modification does not occur in isolation and is often part of a complex series of hydroxylation reactions facilitated by enzymes such as prolyl hydroxylase and ascorbate (vitamin C) as a cofactor.
- Pathways
Hydroxyproline is integral to collagen biosynthesis and modification pathways. One significant pathway is collagen fibril biosynthesis where hydroxyproline ensures proper folding and assembly of collagen fibers. In the same pathway proteins like lysyl oxidase also interact with hydroxyproline-modified collagen to facilitate cross-linking endowing the collagen fibers with added tensile strength. Another important pathway is the post-translational modification cascade involved in protein maturation where hydroxyproline plays a role alongside other hydroxylated amino acids.
- Associated diseases and disorders
Hydroxyproline levels are implicated in conditions like osteogenesis imperfecta and scurvy. Osteogenesis imperfecta caused by a defect in collagen type I features disrupted hydroxyproline incorporation leading to brittle bones. Hydroxyproline is also connected to scurvy where vitamin C deficiency impairs proline hydroxylation resulting in weakened collagen structures. In these cases monitoring hydroxyproline can assist in assessing disease progression and therapeutic outcomes highlighting its relevance alongside proteins such as collagen types I and III.
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2 product images
Functional Studies - Hydroxyproline Assay Kit (Colorimetric) (ab222941)
Functional Studies - Hydroxyproline Assay Kit (Colorimetric) (ab222941)
Typical Hydroxyproline standard calibration curve.
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Product protocols
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