Human OC/BGP(Osteocalcin) ELISA Kit

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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H1343

      Synonyms:BGLAP, BGP, OC, OCN, bone gamma-carboxyglutamate protein, Osteocalcin

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Human

      Detection Range: 1.25~80 ng/mL

      Sensitivity: 0.75 ng/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human OC/BGP concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human OC/BGP. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human OC/BGP and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human OC/BGP, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human OC/BGP. You can calculate the concentration of Human OC/BGP in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 1.25—80 ng/mL
      Sensitivity 0.75 ng/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human OC/BGP in samples. No significant cross-reactivity or interference between Human OC/BGP and analogues was observed.

      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration
      (ng/mL)
      O.D Average Corrected
      80 2.402
      2.442
      2.422 2.357
      40 1.519
      1.543
      1.531 1.466
      20 0.927
      0.895
      0.911 0.846
      10 0.452
      0.482
      0.467 0.402
      5 0.268
      0.266
      0.267 0.202
      2.5 0.175
      0.149
      0.162 0.097
      1.25 0.11
      0.12
      0.115 0.05
      0 0.06
      0.07
      0.065 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human OC/BGP were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human OC/BGP were tested on 3 different plates, 20 replicates in each plate.

      Intra-assay Precision Inter-assay Precision
      Sample 1 2 3 1 2 3
      n 20 20 20 20 20 20
      Mean (ng/mL) 3.70 9.20 32.90 3.80 9.50 36.10
      Standard deviation 0.20 0.40 1.60 0.30 0.50 1.30
      C V (%) 5.41 4.35 4.86 7.89 5.26 3.60

      Recovery

      The recovery of Human OC/BGP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

      Sample Type Range (%) Average Recovery (%)
      Serum (n=5) 92-109 99
      EDTA plasma (n=5) 93-109 99
      Cell culture media (n=5) 85-97 90

      Linearity

      Samples were spiked with high concentrations of Human OC/BGP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 87-102 91-108 91-105
      Average (%) 94 98 97
      1:4 Range (%) 94-109 80-95 95-108
      Average (%) 101 87 102
      1:8 Range (%) 101-117 83-95 98-112
      Average (%) 107 88 104
      1:16 Range (%) 99-113 83-97 90-102
      Average (%) 107 89 97

      Kit components & Storage

      An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 6 months
      Biotinylated Detection Ab Diluent 1 vial, 14 mL
      HRP Conjugate Diluent 1 vial, 14 mL
      Concentrated Wash Buffer (25×) 1 vial, 30 mL
      Substrate Reagent 1 vial, 10 mL 4℃(shading light)
      Stop Solution 1 vial, 10 mL 4℃
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37℃
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.

      2.Remove the liquid.

      3.Add 100 μL Biotinylated Detection Ab. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.

      4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.

      5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

      6.Add 50 μL Stop Solution.

      7.Read at 450 nm immediately. Calculation of results.

      Citations

      1. Publication: Ma W H, Liu Y J, Wang W, et al. Neuropeptide Y, Substance P, And Human Bone Morphogenetic Protein 2 Stimulate Human Osteoblast Osteogenic Activity By Enhancing Gap Junction Intercellular Communication[J]. Brazilian Journal of Medical and Biological Research, 2015, 48(4): 299-307.
        Sample Type: culture supernatant
      2. Publication: He L Y, Zhang X M, Liu B, et al. Effect Of Magnesium Ion On Human Osteoblast Activity[J]. Brazilian Journal of Medical and Biological Research, 2016.
        Sample Type: culture supernatant
      3. Publication: Han H S, Ju F, Geng S. In Vivo And In Vitro Effects Of PTH1-34 On Osteogenic And Adipogenic Differentiation Of Human Bone Marrow-Derived Mesenchymal Stem Cells Through Regulating MicroRNA-155[J]. Journal of Cellular Biochemistry, 2017.
        Sample Type: cell lysis
      4. Publication: Chen H, Wang Y, Dai H, et al. Bone and plasma citrate is reduced in osteoporosis[J]. Bone, 2018.
        Sample Type: Human
      5. Publication: Tamburaci S, Tihminlioglu F. Biosilica incorporated 3D porous scaffolds for bone tissue engineering applications[J]. Materials Science and Engineering: C, 2018, 91: 274-291.
        Sample Type: Human
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