Hydroxyproline ELISA Kit

Cat# OKEH02551

Size : 96Wells

Brand : Aviva Systems Biology

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Phone : +1 850 650 7790
Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
COA DatasheetPrintable COA datasheet for OKEH02551
Product Info
Predicted Species ReactivityAll Species
ApplicationEnzyme-linked Immunosorbent assay-Competitive
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva Systems Biology Hydroxyproline ELISA Kit (OKEH02551) is based on a competitive enzyme immunoassay technique. The microtiter well-plate in this kit has been pre-coated with an anti-Hydroxyproline antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated Hydroxyproline and incubated. The Hydroxyproline found in the sample or standards competes with the biotinylated Hydroxyproline for limited binding sites on the immobilized anti-Hydroxyproline antibody. Excess unbound biotinylated Hydroxyproline and sample or standard Hydroxyproline is washed from the plate. Avidin-HRP conjugate is added, incubated and washed. An enzymatic reaction is then produced through the addition of TMB substrate which is catalyzed by the immobilized HRP to generate a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration is measured by reading the absorbance at 450 nm which is quantitatively proportional to the amount of biotinylated Hydroxyproline captured in the well and inversely proportional to the amount of Hydroxyproline which was contained in the sample or standard.
ELISA Kit Range62.5-4000nmol/L
ELISA Kit RecoveryMean recovery when spiking into Serum and Plasma = 97%
ELISA Kit ReproducibilityMean Intra-assay CV%: < 4.3% (n = 20)
Mean Inter-assay CV%: < 7.5% (n = 20)
ELISA Kit Component
ComponentAmount
Anti-Hydroxyproline Microplate96 Wells (12 x 8 Well strips)
Hydroxyproline Lyophilized Standard2 x 4,000 nmol
Sample Diluent1 x 20 mL
100X Hydroxyproline-Biotin Complex1 x 60 uL
100X Avidin-HRP Conjugate1 x 120 uL
Biotin Complex Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
25X Wash Buffer1 x 30 mL
TMB Substrate1 x 10 mL
Stop Solution1 x 10 mL
Additional InformationTarget Abbr: HYP
Target Name: Hydroxyproline
::Pubchem: 825
::Chemical formula: C5H9NO3
Reconstitution and StorageStore as indicated in product manual.
Sample TypeSerum, plasma, tissue homogenates, cell culture supernatants and other biological fluids
Sensitivity31.25nmol/L
SpecificityNatural and recombinant General Hydroxyproline
Assay InfoAssay Methodology: Quantitative Competitive ELISA
Alias SymbolsHYP, Hydroxyproline, HYP,
Protein NameHydroxyproline
Description of TargetHydroxyproline 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.