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Rat OPG(Osteoprotegerin) ELISA Kit

Cat.No.:E-EL-R3005
Reactivity: Rat

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat OPG in samples. No significant cross-reactivity or interference between Rat OPG and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Rat OPG 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 OPG. 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 OPG 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 OPG, 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 OPG. You can calculate the concentration of Rat OPG in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 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	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat OPG were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat OPG 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 (pg/mL)	45.96	124.69	487.62	44.44	104.83	417.20
Standard deviation	2.56	6.00	26.62	2.29	4.73	17.31
CV (%)	5.57	4.81	5.46	5.15	4.51	4.15

Recovery

The recovery of Rat OPG 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=8)	99-112	105
EDTA plasma (n=8)	93-106	99
Cell culture media (n=8)	90-100	93

Linearity

Samples were spiked with high concentrations of Rat OPG and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	TNFRSF11B , OCIF, TR1
Research Area	Cancer, Cardiovascular, Metabolism, Immunology, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Frontiers in Endocrinology (2021) IF: 5.555

Chondroitin Sulfate Alleviates Diabetic Osteoporosis and Repairs Bone Microstructure via Anti-Oxidation, Anti-Inflammation, and Regulating Bone Metabolism

DOI: 10.3389/fendo.2021.759843
PMID: 34777254	Sample: serum

Frontiers in Pharmacology (2021) IF: 5.811

Salvianolate Ameliorates Osteopenia and Improves Bone Quality in Prednisone-Treated Rheumatoid Arthritis Rats by Regulating RANKL/RANK/OPG Signaling

DOI: 10.3389/fphar.2021.710169
PMID: 34552485	Sample: serum

Nutrients (2018) IF: 4.196

The Effects of Quassinoid-Rich Eurycoma longifolia Extract on Bone Turnover and Histomorphometry Indices in the Androgen-Deficient Osteoporosis Rat Model

DOI: 10.3390/nu10070799
	Sample: Serum

STEM CELLS AND DEVELOPMENT (2020) IF: 3.153

Overloaded orthopedic force induces condylar subchondral bone absorption by stimulating Mesenchymal Stem Cells differentiating into osteoclasts via mTOR-regulated RANKL/OPG secretion in osteoblasts

DOI: 10.1089/scd.2020.0163
	Sample: Tissue homogenate

INFLAMMOPHARMACOLOGY (2018) IF: 3.304

) protected against osteoarthritis by mitigating inflammation and cartilage degradation: a preclinical study

DOI: 10.1007/s10787-017-0432-2
	Sample: serum

PHARMACEUTICAL BIOLOGY (2021) IF: 3.503

Guizhi-Shaoyao-Zhimu decoction attenuates bone erosion in rats that have collagen-induced arthritis via modulating NF‐κB signalling to suppress osteoclastogenesis

DOI: 10.1080/13880209.2021.1876100
	Sample: serum

Veterinary Sciences (2022) IF: 2.518

Histological and Biochemical Analysis after Posterior Mandibular Displacement in Rats

DOI: 10.3390/vetsci9110625
	Sample: serum

MENOPAUSE-THE JOURNAL OF THE NORTH AMERICAN MENOPAUSE SOCIETY (2018) IF: 2.673

Orthosiphon stamineus (Misai Kucing) ameliorated postmenopausal osteoporosis in rat model

DOI: 10.1097/GME.0000000000000980
	Sample: serum

PeerJ (2021) IF: 2.984

Effects of maternal taurine supplementation on maternal dietary intake, plasma metabolites and fetal growth and development in cafeteria diet fed rats

DOI: 10.7717/peerj.11547
	Sample: plasma

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