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Rat OC/BGP(Osteocalcin) ELISA Kit

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Price:   $495
Cat Number:  E-EL-R0243
Lead Time:   7~10 days

Reactivity:  Rat

Detection Range:  0.78~50 ng/mL

Sensitivity:  0.47 ng/mL

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Intended use

This ELISA kit applies to the in vitro quantitative determination of Rat 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 Rat 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 Rat 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 Rat 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 Rat OC/BGP. You can calculate the concentration of Rat OC/BGP in the samples by comparing the OD of the samples to the standard curve.

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Reactivity Rat
Detection Method Colormetric
Detection Range 0.78—50 ng/mL
Sensitivity 0.47 ng/mL
Sample Volume Required Per Well 100μL
Sample Type serum, plasma and other biological fluids

Specificity

This kit recognizes Rat OC/BGP in samples. No significant cross-reactivity or interference between Rat 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
50 2.508
2.566
2.537 2.471
25 1.61
1.662
1.636 1.57
12.5 0.92
0.916
0.918 0.852
6.25 0.493
0.503
0.498 0.432
3.13 0.262
0.24
0.251 0.185
1.56 0.179
0.159
0.169 0.103
0.78 0.116
0.122
0.119 0.053
0 0.062
0.07
0.066 --

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat 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 Rat 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) 2.70 4.70 20.10 2.80 5.10 19.80
Standard deviation 0.10 0.30 1.00 0.20 0.20 0.80
C V (%) 3.70 6.38 4.98 7.14 3.92 4.04

Recovery

The recovery of Rat 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) 85-98 91
EDTA plasma (n=5) 86-96 91
Cell culture media (n=5) 95-109 102

Linearity

Samples were spiked with high concentrations of Rat 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 (%) 86-99 100-115 89-102
Average (%) 91 106 95
1:4 Range (%) 99-113 83-93 94-107
Average (%) 107 88 101
1:8 Range (%) 98-113 83-98 91-107
Average (%) 105 90 99
1:16 Range (%) 101-114 81-93 98-111
Average (%) 107 88 104

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: Kim B, Lee S H, Song S J, et al. Protective Effects of Melon Extracts on Bone Strength, Mineralization, and Metabolism in Rats with Ovariectomy-Induced Osteoporosis[J]. Antioxidants, 2019, 8(8): 306.
    Sample Type: Plasma
  2. Publication: Yang X, Jiang T, Wang Y, et al. The Role and Mechanism of SIRT1 in Resveratrol-regulated osteoblast Autophagy in osteoporosis Rats[J]. Scientific reports, 2019, 9(1): 1-15.
    Sample Type: Serum
  3. Publication: Deng Y, Kang W, Zhao J, et al. Osteoprotective Effect Of Echinocystic Acid, A Triterpone Component From Eclipta Prostrata, In Ovariectomy-Induced Osteoporotic Rats[J]. Plos One, 2015.
    Sample Type: Serum
  4. Publication: Ibrahim K G, Wright H L, Chivandi E, et al. The potential developmental programming effect of oral curcumin on the bone health and plasma total osteocalcin of male and female rats fed a high-fructose diet during suckling and post weaning[J]. General physiology and biophysics, 2019, 38(5): 435-444.
    Sample Type: Plasma
  5. Publication: Niu Y B, Kong X H, Li Y H, et al. Radix Dipsaci Total Saponins Stimulate MC3T3-E1 Cell Differentiation Via The Bone Morphogenetic Protein-2/MAPK/Smad-Dependent Runx2 Pathway[J]. Molecular Medicine Reports, 2015, 11: 4468-4472.
    Sample Type: Cell lysate
  6. Publication: Yang X, He H, Zhou Y, et al. Pulsed Electromagnetic Field At Different Stages Of Knee Osteoarthritis In Rats Induced By Low-Dose Monosodium Iodoacetate: Effect On Subchondral Trabecular Bone Microarchitecture And Cartilage Degradation[J]. Bioelectromagnetics, 2017, 38(3): 227-238.
    Sample Type: Serum,Urine
  7. Publication: Zheng X, Wu G, Nie Y, et al. Electroacupuncture At The Governor Vessel And Bladder Meridian Acupoints Improves Postmenopausal Osteoporosis Through Osteoprotegerin/RANKL/RANK And Wnt/β-Catenin Signaling Pathways[J]. Experimental & Therapeutic Medicine, 2015, 10: 541-548.
    Sample Type: Serum
  8. Publication: Tantowi N A C A, Mohamed S, Lau S F, et al. Comparison of diclofenac with apigenin-glycosides rich Clinacanthus nutans extract for amending inflammation and catabolic protease regulations in osteoporotic-osteoarthritis rat model[J]. Daru: journal of Faculty of Pharmacy, Tehran University of Medical Sciences, 2020.
    Sample Type: Serum
  9. Publication: Yuliyanasari N, Mastutik G, Putra S T. THE ELEVATION OF OSTEOBLAST ACTIVITY IN RAT BONE MARROW MESENCHYMAL STEM CELLS IN OSTEOGENIC MEDIUM EXPOSED WITH MELATONIN IN PHYSIOLOGICAL DOSES[J]. Folia Medica Indonesiana, 2017, 53(1): 41-48.
    Sample Type: Cell lysate
  10. Publication: Costa N, Gomes A, Silva C, et al. Symbiotic Effects of Kefir and Yacon Flour (Smallanthus sonchifolius) on Bone Remodeling in Rats[J]. Current Developments in Nutrition, 2020, 4(Supplement_2): 384-384.
    Sample Type: Urine,Feces
  11. Publication: Nurdiana N, Mariati N, Noorhamdani N, et al. Effects of Labisia pumila on bone turnover markers and OPG/RANKL system in a rat model of post-menopausal osteoporosis[J]. Clinical Nutrition Experimental, 2018, 20: 41-47.
    Sample Type: Serum,Urine
  12. Publication: Chinnappan S M, George A, Ashok G, et al. Effect of herbal extract Eurycoma longifolia (Physta) on female reproductive hormones and bone biochemical markers: an ovariectomised rat model study[J]. BMC Complementary Medicine and Therapies, 2020, 20(1): 1-9.
    Sample Type: Serum
  13. Publication: Sananta P, Jonatan A, Ernanda S A, et al. Effects of nicotine on markers of bone turnover in ovariectomized rats[J]. The Pan African Medical Journal, 2019, 33.
    Sample Type: Serum
  14. Publication: Giaze T R, Shuid A N, Soelaiman I N, et al. Bone turnover and oxidative stress markers in estrogendeficient rats treated with Marantodes pumilum leaves and roots: A comparative study[J]. Tropical Journal of Pharmaceutical Research, 2018, 17(4): 611-617.
    Sample Type: Blood,Tissue homogenate
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