25-OH Vitamin D ELISA Kit
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- Target See all 25-OH Vitamin D products
- 25-OH Vitamin D
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Reactivity
- Chemical
- Detection Method
- Colorimetric
- Application
- ELISA
- Purpose
- 25-OH Vitamin D ELISA assay kit is designed for the serological determination of the vitamin D concentration in the human organism
- Sample Type
- Serum, Plasma
- Sensitivity
- 1.9 ng/mL
- Characteristics
- Types of vitamin D that are differentiated are vitamin D2 (ergocalciferol) that is contained in plant food (mushrooms, avocado) and vitamin D3 (cholecalciferol) that is produced from 7-dehydrocholesterol in the skin under ultra-violet irradiation or found in animal food or products (sea fish, egg yolk, butter) [1, 2, 3, 4]. These two forms of vitamin D, which are not yet biologically active, are bound by a protein called VDBP (vitamin D binding protein) in the bloodstream, then metabolised in the liver and converted into 25-OH vitamin D2 (calcidiol) and 25-OH vitamin D3 (calcitriol), respectively, which are storage forms of the vitamin with little activity [1]. In contrast to other commercially available tests, the Eagle Biosciences 25-OH Vitamin D ELISA assay kit uses a newly designed monoclonal antibody which is specific for both vitamin D2 and vitamin D3 at 100 % specificity. This is necessary because sometimes vitamin D2 instead of D3 is used in therapy [5, 6, 7].
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- Application Notes
- Optimal working dilution should be determined by the investigator.
- Sample Volume
- 10 μL
- Assay Time
- 2 - 3 h
- Protocol
- The new 25-OH Vitamin D ELISA assay test kit is designed for the determination of 25-OH Vitamin D in human serum or plasma samples. In the first analysis step, the calibrators and patient samples are diluted with biotin-labelled 25-OH vitamin D and added to microplate wells coated with monoclonal anti-25-OH Vitamin D antibodies. During the incubation an unknown amount of 25-OH Vitamin D in the patient sample and a known amount of biotin-labelled 25-OH vitamin D compete for the antibody binding sites in the microplate wells plate. Unbound 25-OH vitamin D is removed by washing. For the detection of bound biotin-labelled 25-OH vitamin D, a second incubation is performed using peroxidase-labelled streptavidin. In a third incubation using the peroxidase substrate tetramethylbenzidine (TMB) the bound peroxidase promotes a colour reaction. The colour intensity is inversely proportional to the 25-OH vitamin D concentration.
- Restrictions
- For Research Use only
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- Storage
- 4 °C
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- Target See all 25-OH Vitamin D products
- 25-OH Vitamin D
- Abstract
- 25-OH Vitamin D Products
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