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DHVD3(1,25-Dihydroxyvitamin D3) ELISA Kit

Cat.No.:E-EL-0016
Reactivity: Universal

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	7.81-500 pg/mL
Sensitivity	4.69 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes DHVD3 in samples. No significant cross-reactivity or interference between DHVD3 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 DHVD3 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 DHVD3. During the reaction, DHVD3 in the sample or standard competes with a fixed amount of DHVD3 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to DHVD3. 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 DHVD3 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 DHVD3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level DHVD3 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)	26.85	42.69	171.01	25.28	43.72	154.82
Standard deviation	1.47	1.93	8.12	1.29	2.28	4.92
CV (%)	5.47	4.52	4.75	5.10	5.22	3.18

Recovery

The recovery of DHVD3 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)	92-105	98
EDTA plasma(n=8)	88-101	94
Cell culture media(n=8)	88-103	94

Target Information

Research Area

Metabolism, Developmental biology

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

Food & Function (2022) IF: 5.396
Diosmin, a citrus fruit-derived phlebotonic bioflavonoid protects rats from chronic kidney disease-induced loss of bone mass and strength without deteriorating the renal function

DOI: 10.1039/D1FO03867B

PMID: 35119062
Sample: serum
Frontiers in Psychiatry (2022) IF: 5.435
Efficacy of a novel proprietary dietary supplement (TRI 360TM) on psychological symptoms and stress-related quality of life in adult subjects: A randomized controlled clinical trial

DOI: 10.3389/fpsyt.2022.919284

PMID: 36032243
Sample: serum
POULTRY SCIENCE (2019) IF: 2.027
Effects of dietary organic minerals, fish oil, and hydrolyzed collagen on growth performance and tibia characteristics of broiler chickens

DOI: 10.3382/ps/pez427

PMID: 31392338
Sample: Serum
JOURNAL OF INTERNATIONAL MEDICAL RESEARCH (2020) IF: 1.287
Bacillus Calmette-Guérin (BCG) stimulates changes in dendritic cell surface marker expression in vitamin D-deficient mice:

DOI: 10.1177/0300060519896892

Sample: Serum
Journal of General and Family Medicine (2023)
The role of biofield energy treatment on psychological symptoms, mental health disorders, and stress-related quality of life in adult subjects: A randomized controlled clinical trial

DOI: 10.1002/jgf2.606

Sample: serum
International Journal of Clinical and Experimental Pathology (2019) IF: 0.205
miR-193-3p ameliorates bone resorption in ovariectomized mice by blocking NFATc1 signaling

PMID: 31933803
Sample: serum

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