Serum, plasma, tissue homogenates, cell culture supernatants, other biological fluids
Species Reactivity
Human
Detection Method
cELISA
Intended Use
For the quantitative measurement of Hydroxyproline in serum, plasma, tissue homogenates, cell culture supernatants and other biological fluids.
Contents of Kit
1.Anti-Hydroxyproline Microplate 96 Wells, (12 x 8 Well strips) 2.Hydroxyproline Lyophilized Standard, 2 x 8,000 nmol 3.Sample Diluent, 1 x 20 mL 4.100X Hydroxyproline-Biotin Complex, 1 x 60 μL 5.100X Avidin-HRP Conjugate, 1 x 120 μL 6.Biotin Complex Diluent, 1 x 12 mL 7.Conjugate Diluent, 1 x 12 mL Storage Conditions: -20°C for 6 months 8.25X Wash Buffer, 1 x 30 mL 9.TMB Substrate, 1 x 10 mL 10.Stop Solution, 1 x 10 mL Storage Conditions: 4°C for 6 months
Storage
Open kit immediately upon receipt. Store components at -20°C or 4°C for 6 months or until expiration date. Avoid any freeze/thaw cycles.
Detection Range
62.5 - 4,000 nmol/L
Detection Limit
< 31.25 nmol/L (Derived by linear regression of OD450 of the Mean Blank + 2xSD)
General Description
Hydroxyproline is a major component of the protein collagen. Hydroxyproline and proline play key roles for collagen stability. They permit the sharp twisting of the collagen helix. Hydroxyproline is found in few proteins other than collagen. For this reason, hydroxyproline content has been used as an indicator to determine collagen and/or gelatin amount. Hydroxyproline is produced by hydroxylation of the amino acid proline by the enzyme prolyl hydroxylase following protein synthesis (as a post-translational modification). The enzyme catalysed reaction takes place in the lumen of the endoplasmic reticulum. Although it is not directly incorporated into proteins, hydroxyproline comprises roughly 4% of all amino acids found in animal tissue, more than seven amino acids which are directly incorporated.
Standard Curve
This standard curve is for demonstration purposes only. An assay specific standard curve should be performed with each assay.
Citations
Publication ()
Have you cited DEIA3605 in a publication? Let us know and earn a reward for your research.
Background
Hydroxyproline (Hyp) is a hydroxylated derivative of proline that is formed during the translation and modification processes of proteins, making it widely distributed in proteins and particularly abundant in collagen. Collagen, a major structural protein found in tissues such as skin, bone and muscle, is the major component of connective tissue in the body. The metabolism of hydroxyproline is essential for maintaining normal physiological functions. Hydroxylation of proline typically occurs in the endoplasmic reticulum and is catalyzed by specific enzymes. Two isomeric forms of hydroxyproline, trans-4-hydroxy-L-proline and trans-3-hydroxy-L-proline, contribute to the triple helical structure of collagen, ensuring its strength and structural stability in tissues. This role makes hydroxyproline essential in the composition of collagen.
Hydroxyproline finds significant applications in basic research and therapeutic developments. Diseases such as osteoporosis, rheumatoid arthritis, atherosclerosis, aortic aneurysm, and vitreous humor diseases are linked to abnormalities in collagen structure or function. Therefore, hydroxyproline serves as a biomarker for evaluating treatment efficacy and developing novel therapeutic strategies for these conditions. It also supports maintaining collagen structure and function in skin, aiding conditions like systemic lupus erythematosus and slowing down skin aging. Furthermore, due to its role in protein structure, hydroxyproline is utilized in developing artificial tissues as scaffold proteins or materials for tissue repair, contributing significantly to regenerative medicine.
Hydroxyproline can also help enhance agricultural efficiency. As a soil conditioner, hydroxyproline improves soil structure, water retention, and fertility. The application of hydroxyproline to plants stimulates growth, enhances drought resistance, protects against oxidative stress, and promotes plant health. As a feed additive for livestock, hydroxyproline not only maintain the health of their skin, bones, and joints., but it also protects the intestines by reducing inflammatory reactions. Feeding fishmeal with hydroxyproline can improve the hardness and texture of shrimp, fish, and shellfish meat.
Creative Diagnostics products are for RESEARCH USE ONLY, please make sure your review is research based.
Required fields are marked with *
Terms and conditions:
We will select high-quality review customers and offer a $30 coupon for your next purchase.
All product reviews must be submitted in the English language.
Creative Diagnostics will not share any personal information of applicants, and all information will be treated with strict confidentiality and will not be sold or disclosed to a third party.
References
Derivatization-based magnetic dummy molecularly imprinted polymers integrated with 4-plex stable isotope labeling derivatization strategy for specific and rapid determination of L-hydroxyproline in human serum
The specific determination of L-hydroxyproline (Hyp) can serve as a potential indicator for early clinical diagnosis of liver fibrosis. In this work, an integrated strategy based on 4-plex stable isotope labeling derivatization combined with dummy magnetic molecularly imprinted polymers (QSILD-DMMIPs) was developed for specific extraction and rapid determination of Hyp in human serum by ultra high performance liquid chromatography tandem mass spectrometry. A new series of QSILD reagents d(0)/d(1)/d(2)/d(3)-6-N-methyl-rhodamine 6G-N-hydroxysuccinimidyl formate (d(0)/d(1)/d(2)/d(3)-MRSF) were designed, synthesized and applied for the high-throughput labeling of Hyp in serum samples. The structural analogue derivative of Hyp with 6-N-ethyl-rhodamine 6G-N-hydroxysuccinimidyl formate (ERSF-Hyp) was synthesized and used as a novel dummy template to prepare DMMIPs. The DMMIPs were well characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), fourier transform infrared spectroscopy (FTIR), Brunner Emmet Teller (BET) measurements, thermogravimetric analysis (TGA), X-ray diffraction (XRD), zeta potential and adsorption experiments. All d(0)/d(1)/d(2)/d(3)-MRSF-Hyp derivatives were conveniently and specifically adsorbed by DMMIPs in magnetic dispersive solid phase extraction procedure before injection. Method validation results including linearity (0.2-100 ng mL(-1)), limits of detection and quantitation (0.05 and 0.2 ng mL(-1)), accuracy, precision, stability, matrix effect and derivatization efficiency were satisfactory. The analytical performances benefited from efficient integration of QSILD and specific DMMIPs extraction. The proposed strategy was successfully applied for Hyp determination in human serum of liver fibrosis patients and healthy controls, which was of great significance to early diagnosis. (C) 2020 Elsevier B.V. All rights reserved.
Generation of bioactive prolyl-hydroxyproline (Pro-Hyp) by oral administration of collagen hydrolysate and degradation of endogenous collagen
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
Hydroxyproline-containing di- and tripeptides increase in human blood plasma after ingestion of collagen hydrolysate in a dose-dependent manner. Prolyl-hydroxyproline (Pro-Hyp) and hydroxyprolyl-glycine (Pro-Gly) are the main food-derived collagen peptides present in human blood plasma. Pro-Hyp increases the number of fibroblasts migrating from the mouse skin and enhances the growth of fibroblasts attached to the collagen gel. Pro-Hyp is also generated from endogenous collagen in the granulation tissue in mice skin and ear with dermatitis, while it does not increase in the normal tissues of the same animal. However, Hyp-Gly does not increase in all tissues. These facts indicate that Pro-Hyp is generated by the degradation of endogenous collagen to activate cells involved in tissue reconstruction. Orally administered Pro-Hyp is deposited in animal organs even after clearance of Pro-Hyp from blood. Thus, the endogenous bioactive peptide can be provided by ingestion of collagen hydrolysate.
COA
Publication 
Loading ......
