United States set
Menu Shopping cart $0 Search
Distributed product

25-OH-Vitamin-D ELISA

  • Regulatory status:RUO
  • Type:Competitive ELISA
  • Species:Human
This product is not available in United States!
Cat. No. Size Price

EA300/96 96-wells
PubMed Product Details
Technical Data

Cat # changed from REA300/96 to EA300/96


Competitive ELISA


The new ELISA test kit is designed for the in vitro determination of 25-OH Vitamin D in human serum or plasma samples.


Serum, Plasma-EDTA, Plasma-Heparin, Plasma-Citrate

Sample Requirements

20 µl


On blue ice packs. Upon receipt, store the product at the temperature recommended below.


Store the complete kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box).

Calibration Curve

Calibration Range

0 – 120 ng/ml

Limit of Detection

1.6 ng/ml

Intra-assay (Within-Run)

n = 40; CV = 5.0 %

Inter-assay (Run-to-Run)

n = 10; CV = 7.8 %

Dilution Linearity




  • RUO
  • standards 1-6: 0-120 ng/ml
  • limit of detection 1.6 ng/ml
  • Material of animal origin used in the preparation of the kit have been obtained from certified healthy animals but these materials should be handled as potentially infectious

Research topic

Bone and cartilage metabolism


The 25-OH Vitamin D ELISA is designed for the serological determination of the Vitamin D concentration in the human organism. 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 D3 (calcidiol) and subsequently to the biological active form 1,25(OH)2 Vitamin D3 (calcitriol) in the kidney [1]. In contrast to other commercially available tests, the ELISA uses a newly designed monoclonal antibody which is equally specific for both forms of the vitamin. This is necessary because sometimes Vitamin D2 instead of D3 is used in therapy [5, 6, 7]. The new ELISA test kit is designed for the in vitro determination of 25-OH Vitamin D in human serum or plasma samples. In the first analysis step, the calibrators and 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 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.

Summary References (7)

References to 25-OH-Vitamin-D

  • Holick MF, Chen TC. Vitamin D deficiency: a worldwide problem with healthconsequences. Am J Clin Nutr. 2008 Apr;87(4):1080S-6S. Review. PubMed PMID:18400738. See more on PubMed
  • Hollis BW. Editorial: The determination of circulating 25-hydroxyvitamin D: noeasy task. J Clin Endocrinol Metab. 2004 Jul;89(7):3149-51. PubMed PMID:15240585. See more on PubMed
  • Lips P. Vitamin D physiology. Prog Biophys Mol Biol. 2006 Sep;92(1):4-8. Epub 2006 Feb 28. Review. PubMed PMID: 16563471. See more on PubMed
  • Macdonald HM, Mavroeidi A, Fraser WD, Darling AL, Black AJ, Aucott L, O'Neill F, Hart K, Berry JL, Lanham-New SA, Reid DM. Sunlight and dietary contributionsto the seasonal vitamin D status of cohorts of healthy postmenopausal womenliving at northerly latitudes: a major cause for concern? Osteoporos Int. 2011Sep;22(9):2461-72. doi: 10.1007/s00198-010-1467-z. Epub 2010 Nov 18. Erratum in: Osteoporos Int. 2011 Sep;22(9):2473-4. PubMed PMID: 21085934. See more on PubMed
  • Mavroeidi A, O'Neill F, Lee PA, Darling AL, Fraser WD, Berry JL, Lee WT, Reid DM, Lanham-New SA, Macdonald HM. Seasonal 25-hydroxyvitamin D changes in British postmenopausal women at 57 degrees N and 51 degrees N: a longitudinal study. JSteroid Biochem Mol Biol. 2010 Jul;121(1-2):459-61. doi:10.1016/j.jsbmb.2010.03.038. Epub 2010 Mar 17. PubMed PMID: 20302933. See more on PubMed
  • Snellman G, Melhus H, Gedeborg R, Byberg L, Berglund L, Wernroth L,Michaëlsson K. Determining vitamin D status: a comparison between commerciallyavailable assays. PLoS One. 2010 Jul 13;5(7):e11555. doi:10.1371/journal.pone.0011555. Erratum in: PLoS One. 2010;5(9). doi:10.1371/annotation/23307aa4-726e-4f11-86c0-8a292be33517. PubMed PMID: 20644628;PubMed Central PMCID: PMC2903481. See more on PubMed
  • Tsur A, Metzger M, Dresner-Pollak R. Effect of different dress style onvitamin D level in healthy young Orthodox and ultra-Orthodox students in Israel. Osteoporos Int. 2011 Nov;22(11):2895-8. doi: 10.1007/s00198-010-1492-y. Epub 2010Nov 26. PubMed PMID: 21110005. See more on PubMed
Related Products Docs