The IDS 1,25-Dihydroxy Vitamin D EIA kit is a complete assay system for the purification of 1,25D in patient samples by immunoextraction followed by quantitation by EIA. Patient samples are delipidated and 1,25D extracted from potential cross-reactants by incubation for 90 minutes with a highly specific solid phase monoclonal anti-1,25D. The immunoextraction gel is then washed and purified 1,25D eluted directly into glass assay tubes. Reconstituted eluates and calibrators are incubated overnight with a highly specific sheep anti-1,25D. Then a portion of this is incubated for 90 minutes with shaking in microplate wells which are coated with a specific anti-sheep antibody. 1,25D linked to biotin is then added and the plate shaken for a further 60 minutes before aspiration and washing. Enzyme (horseradish peroxidase) labelled avidin is added and binds selectively to complexed biotin and, following a further wash step, colour is developed using a chromogenic substrate (TMB). The absorbance of the stopped reaction mixtures are read in a microtitre plate reader, colour intensity developed being inversely proportional to the concentration of 1,25D.
Size
96 wells
Concentration
n/a
Applications
colorimetric ELISA or EIA
Other Names
1,25-dihydroxyvitamin D; 1,25D; 1,25OH D kit, assay, immunoassay