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Human DBP(Vitamin D Binding Protein) ELISA Kit

Citations(6)Uniprot : P02774

Cat.No.:E-EL-H1604
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	3.91-250 ng/mL
Sensitivity	2.35 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human DBP in samples. No significant cross-reactivity or interference between Human DBP 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 DBP 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 DBP. 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 DBP 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 DBP, 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 DBP. You can calculate the concentration of Human DBP 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 DBP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human DBP 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)	12.10	23.12	91.12	12.04	22.67	95.55
Standard deviation	0.70	1.17	3.78	0.69	1.09	5.17
CV (%)	5.79	5.06	4.15	5.73	4.81	5.41

Recovery

The recovery of Human DBP 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)	87-98	92
EDTA plasma (n=8)	89-103	96
Cell culture media (n=8)	86-99	92

Linearity

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

Target Information

Database Links	SwissProt: P02774
Synonyms	Vitamin D-binding protein,gc-globulin ,GC, DBP, DBP/GC, GRD3, VDBG, VDBP
Research Area	Cancer, Metabolism, 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

Cureus (2021)

Associations Between Bioavailable Vitamin D and Remnant Cholesterol in Patients With Type 2 Diabetes Mellitus

DOI: 10.7759/cureus.13248
	Sample: Serum

Turkish Journal of Biochemistry-Turk Biyokimya Dergisi (2021) IF: 0.401

Increased vitamin D binding protein levels are associated with irritable bowel syndrome

DOI: 10.1515/tjb-2020-0305
	Sample: Serum

ARQUIVOS DE NEURO-PSIQUIATRIA (2019) IF: 1.048

Vitamin D, vitamin D binding protein, vitamin D receptor levels and cardiac dysautonomia in patients with multiple sclerosis: a cross-sectional study

DOI: 10.1590/0004-282x20190182
	Sample: Serum

JOURNAL OF ENDOCRINOLOGICAL INVESTIGATION (2019) IF: 3.439

Association of thyroid cancer risk with plasma 25-hydroxyvitamin D and vitamin D binding protein: a case–control study in China

DOI: 10.1007/s40618-019-01167-7
	Sample: Plasma

Clinical Oral Investigations (2023) IF: 3.606

Total, free, and bioavailable 25-hydroxyvitamin D levels in patients with periodontitis (stage III): a case–control study

DOI: 10.1007/s00784-022-04844-9
	Sample: Serum

Nutrients (2021) IF: 5.719

Rapid and Effective Vitamin D Supplementation May Present Better Clinical Outcomes in COVID-19 (SARS-CoV-2) Patients by Altering Serum INOS1, IL1B, IFNg, Cathelicidin-LL37, and ICAM1

DOI: 10.3390/nu13114047
PMID: 34836309	Sample: Serum

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Reviews/Q&A

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Q&A

D*************aSubmitted [ Jan 25 2018 ]

Asked: Hello, Is kit running with monoclonal ab or policlonal ab ?

P*******nSubmitted [ Jan 25 2018 ]

Answered: Dear Dr. Gulsum Feyza, Thanks for your question. The kit is running with both monoclonal Ab and polyclonal Ab. The coated antibody is monoclonal and the detection antibody is polyclonal. Any other questions, please let us know. Or you could email us techsupport@elabscience.com

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