Canine 25-hydroxy vitamin D3 (25 HVD3) ELISA Kit

This assay employs a two-site sandwich ELISA to quantitate 25 HVD3 in samples. An antibody specific for 25 HVD3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any 25 HVD3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for 25 HVD3 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of 25 HVD3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
 

Calcifediol is a prehormone which is produced by hydroxylation of vitamin D3 (cholecalciferol) in the liver. Calcidiol is then converted in the kidneys (by the enzyme 25(OH)D-1
 

Reagents	Quantity	Reagents	Quantity
Assay plate (96 Wells)	1	Instruction manual	1
Standard (lyophilized)	2	Sample Diluent	1 x 20 mL
Biotin-Conjugate (concentrate 100 x)	1 x 120 μL	Biotin-Conjugate Diluent	1 x 12 mL
Streptavidin-HRP  (concentrate 100 x)	1 x 120 μL	Streptavidin-HRP Diluent	1 x 12 mL
Wash Buffer (concentrate 25 x)	1 x 20 mL	Substrate Solution	1 x 10 mL
Stop Solution	1 x 6 mL	Adhesive Films	4

 

 

Sample Type	Number	Recovery range (%)	Average(%)
Serum	10	90-101	96
EDTA plasma	10	89-97	93
Heparin plasma	10	91-99	95

 

 

Sample Type	1：2	1：4	1：8	1：16
Serum	78-89%	81-99%	92-103%	95-105%
EDTA plasma	91-101%	90-98%	93-101%	91-98%
Heparin plasma	92-103%	93-102%	92-99%	91-101%