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VD(Vitamin D) ELISA Kit

Cat.No.:E-EL-0012
Reactivity: Universal

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96T
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96T*5
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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	6.25-400 ng/mL
Sensitivity	3.75 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes VD in samples. No significant cross-reactivity or interference between VD 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 VD concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with VD. During the reaction, VD in the sample or standard competes with a fixed amount of VD on the solid phase supporter for sites on the Biotinylated Detection Ab specific to VD. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of VD in tested samples can be calculated 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℃
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 VD were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level VD 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)	21.62	53.66	157.91	21.41	57.30	152.81
Standard deviation	1.28	2.70	5.98	1.47	2.35	8.34
CV (%)	5.92	5.03	3.79	6.87	4.10	5.46

Recovery

The recovery of VD 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)	96-112	102
EDTA plasma(n=8)	88-99	93
Cell culture media(n=8)	85-97	91

Target Information

Research Area

Metabolism

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

JOURNAL OF NUTRITIONAL BIOCHEMISTRY (2022) IF: 6.117
Vitamin D protects silica particles induced lung injury by promoting macrophage polarization in a KLF4-STAT6 manner

DOI: 10.1016/j.jnutbio.2022.109148

PMID: 36049670
Sample: serum
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY (2021) IF: 6.291
Upregulation of autophagy in M2 macrophage by vitamin D alleviates crystalline silica-induced pulmonary inflammatory damage

DOI: 10.1016/j.ecoenv.2021.112730

PMID: 34478973
Sample: lung
Journal of Medical Biochemistry (2021) IF: 3.402
Low levels of vitamin D were associated with coagulopathy among hospitalized coronavirus disease-19 (COVID-19) patients: A single-centered study in Indonesia

DOI: 10.5937/jomb0-30228

Sample: serum
Tuberkuloz ve Toraks-Tuberculosis and Thorax (2020)
Evaluation of the relationship of serum vitamin D levels in COVID-19 patients with clinical course and prognosis.

DOI: 10.5578/tt.70027

Sample: blood

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Reviews/Q&A

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Reviews
Q&A

r****aSubmitted [ Nov 09 2022 ]

Asked: Can we use this kit for measuring mouse serum samples

adminSubmitted [ Nov 09 2022 ]

Answered: Yes,it is a universal kit and is suitable for mouse serum.

M******************ćSubmitted [ Mar 28 2022 ]

Asked: Does this kit measure levels of 25(OH) vitamin D in saliva samples?

adminSubmitted [ Mar 28 2022 ]

Answered:

D**uSubmitted [ Sep 25 2021 ]

Asked: Hi Does the Vitamin D (VD) ELISA kit ( E-EL-0012) measure calciferol (vitamin D 2 + vitamin D3) levels or 25 (OH) V D ?

adminSubmitted [ Sep 25 2021 ]

Answered: E-EL-0012 measures vitamin D 2 + vitamin D3 levels.

D*nSubmitted [ Sep 25 2021 ]

Asked: Hi Does this kit measure calciferol or 25 (OH) D?

adminSubmitted [ Sep 25 2021 ]

Answered: E-EL-0012 measures vitamin D 2 + vitamin D3 levels. E-EL-0015 measures 25 (OH) Vitamin D3 only.

D******aSubmitted [ Sep 25 2021 ]

Asked: Hi 1. Does this kit measure ( Vitamin D2 + Vitamin D3 ) levels or does it measure 25 (OH) D 3 levels? 2. What is the difference between this ELISA kit (E-EL-0012) and the 25 (OH) D3 ELISA kit (E-EL-0015)?

adminSubmitted [ Sep 25 2021 ]

Answered: E-EL-0012 measures vitamin D 2 + vitamin D3 levels. E-EL-0015 measures 25 (OH) Vitamin D3 only.

S*****eSubmitted [ Sep 25 2021 ]

Asked: Does this kit measure (Vitamin D3 + Vitamin D2) levels or 25 (OH) D3 levels?

adminSubmitted [ Sep 25 2021 ]

Answered: E-EL-0012 measures vitamin D 2 + vitamin D3 levels. E-EL-0015 measures 25 (OH) Vitamin D3 only.

E**eSubmitted [ Mar 26 2019 ]

Asked: Hello, I would like to ask what is evaluated in this test: Free vitamin D, Bioavailable vitamin D or 25(OH)D? Maybe you know the values of this test?

adminSubmitted [ Mar 26 2019 ]

Answered: Hi Egle, This kit detects 25(OH)Vintamin D, the reference values for human serum samples were 20-100 ng/mL.

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