Description
Key facts
Detection method:Colorimetric,
Sample types:Plasma, Serum,
Reacts with:Human,
Assay type:Competitive,
Sensitivity:= 1.98 ng/mL,
Range:0.5 - 1010 ng/mL,
Assay time:1h 30m,
Assay Platform:Pre-coated microplate (12 x 8 well strips)
Product details:
25(OH) Vitamin D ELISA kit ab213966 is a competitive ELISA kit to measure 25-hydroxy vitamin D (vitamin D2 & vitamin D3) in human serum and plasma with a sensitivity of 1.98 ng/mL.
How the assay works
25(OH) Vitamin D ELISA kit ab213966 is a competitive ELISA. There are 2 types of competitive ELISAs:
antigen down
: The sample antigen competes with a reference antigen for binding to a specific amount of labeled antibody. The reference antigen is pre-coated on a multi-well plate. The sample is added to the plate wells with a labeled antibody. Depending on the amount of antigen in the sample, more or fewer free antibodies will be available to bind the reference antigen. This means that the more of the antigen there is in the sample, the less reference antigen will be detected and the weaker the signal will be.
antibody down
: the labeled antigen and the sample antigen (unlabeled) compete for binding to the primary antibody, which is immobilized on the plate. The lower the amount of antigen in the sample, the stronger the signal due to more labelled antigen in the well.
Assay specificity
Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility.
25(OH) Vitamin D ELISA kit ab213966 protocol summary
Add standards or samples to the wells, incubate 2 hrs
Wash, then add 50 µl of Biotinylated Antibody per well. Incubate 1 hr
Wash, then add 50 µl of SP Conjugate per well. Incubate 30 min
Wash, then add 50 µl of Chromogen Substrate per well. Incubate 20 min
Add 50 µl of Stop Solution per well. Read at 450 nm immediately
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
25-OH Vitamin D3 also known as calcidiol or 25-hydroxycholecalciferol serves as a prohormone and the main storage form of vitamin D in the human body. It has a molecular mass of approximately 400.64 g/mol. The liver expresses it where the conversion from vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol) occurs. It circulates in the bloodstream and acts as a reservoir for the active form of vitamin D.
Biological function summary
25-hydroxyvitamin D3 is important to maintaining calcium and phosphate homeostasis. It is not part of a protein complex in its function as a circulatory prohormone. The kidneys convert it into the active form 125-dihydroxyvitamin D3 (calcitriol) which then interacts with the vitamin D receptor (VDR) to modulate gene expression related to calcium metabolism.
Pathways
This vitamin D3 metabolite holds importance in the calcium signaling and bone remodeling pathways. Its conversion to calcitriol aids in the regulation of parathyroid hormone (PTH) ensuring calcium balance in the bloodstream. The pathway includes elements such as the VDR and CYP27B1 which are necessary for its activation and action.
Alterations in 25-OH Vitamin D3 levels influence conditions like osteoporosis and rickets. Both conditions relate to disruptions in calcium homeostasis. Proteins like the VDR and CYP27B1 are involved in these disorders influencing bone density and mineralization through the modulation of calcidiol and calcitriol levels.
