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Human ON(Osteonectin) ELISA Kit

Citations(9)Uniprot : P09486

Cat.No.:E-EL-H1351
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T/24
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human ON in samples. No significant cross-reactivity or interference between Human ON 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 Human ON 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 ON. 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 ON 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 ON, 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 ON. You can calculate the concentration of Human ON 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℃, 12 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), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 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 Human ON were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ON 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)	1.08	1.91	8.12	1.14	1.95	7.33
Standard deviation	0.07	0.11	0.38	0.06	0.10	0.37
CV (%)	6.48	5.76	4.68	5.26	5.13	5.05

Recovery

The recovery of Human ON 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)	89-101	96
EDTA plasma (n=8)	88-102	93
Cell culture media (n=8)	89-104	96

Linearity

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

Target Information

Database Links	SwissProt: P09486
Synonyms	SPARC
Research Area	Cancer, Cell Biology, Cardiovascular, Developmental biology, Signal transduction, Stem cells

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

Biocatalysis and Agricultural Biotechnology (2020)

Prophylactic supplementation of sinapic acid ameliorates zoledronic acid induced changes in osteoblast survival and differentiation

DOI: 10.1016/j.bcab.2020.101700
	Sample: Mg63 Cell

PROCESS BIOCHEMISTRY (2019) IF: 2.883

Zinc chelated morin promotes osteoblast differentiation over its uncomplexed counterpart

DOI: 10.1016/j.procbio.2019.04.008
	Sample: Mg-63 Cells

LIFE SCIENCES (2020) IF: 3.647

Kaempferol-zinc(II) complex synthesis and evaluation of bone formation using zebrafish model

DOI: 10.1016/j.lfs.2020.117993
	Sample: Cell Culture Medium

LIFE SCIENCES (2020) IF: 3.647

Assessment of human ovarian follicular fluid derived mesenchymal stem cells in chitosan/PCL/Zn scaffold for bone tissue regeneration

DOI: 10.1016/j.lfs.2020.118502
PMID: 33031825	Sample: Cell Sample

PEPTIDES (2023) IF: 3.867

Glucagon-like peptide-1 receptor promotes osteoblast differentiation of dental pulp stem cells and bone formation in a zebrafish scale regeneration model

DOI: 10.1016/j.peptides.2023.170974
	Sample: Human Dental Pulp Stem Cells (Hdpscs)

JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY (2022) IF: 3.981

Nano-encapsulation of melatonin into polydiacetylene-phospholipid assembly for sustained-release and enhanced bone formation in zebrafish

DOI: 10.1016/j.jddst.2022.103415
	Sample: Mmscs

EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES (2021) IF: 4.384

Bioactive Zinc(II) complex incorporated PCL/gelatin electrospun nanofiber enhanced bone tissue regeneration

DOI: 10.1016/j.ejps.2021.105768
PMID: 33607242	Sample: Cell Culture Supernatant(Human Mg-63 Cells)

Hepatology Communications (2018) IF: 5.073

Suboptimal Level of Bone-Forming Cells in Advanced Cirrhosis are Associated with Hepatic Osteodystrophy

DOI: 10.1002/hep4.1234
PMID: 30202823	Sample: Plasma,Tissue Slice

CHEMICO-BIOLOGICAL INTERACTIONS (2021) IF: 5.194

Rutin-Zn(II) complex promotes bone formation - A concise assessment in human dental pulp stem cells and zebrafish

DOI: 10.1016/j.cbi.2021.109674
PMID: 34562440	Sample: Hdpsc

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