Calcitriol ELISA Kit

Calcitriol, also called 1,25-dihydroxycholecalciferol or 1,25-dihydroxyvitamin D3, is the hormonally active form of vitamin D with three hydroxyl groups (abbreviated 1,25- (OH) 2D3 or simply 1,25 (OH) 2D) . It increases the level of calcium (Ca2+) in the blood by (1) increasing the uptake of calcium from the gut into the blood, (2) decreasing the transfer of calcium from blood to the urine by the kidney, and (3) increasing the release of calcium into the blood from bone. Calcitriol is produced in the cells of the proximal tubule of the nephron in the kidneys by the action of 25-hydroxyvitamin D3 1-alpha-hydroxylase, a mitochondrial oxygenase and an enzyme which catalyzes the hydroxylation of 25-hydroxycholecalciferol (calcifediol) . The activity of the enzyme is stimulated by PTH. The reaction is an important control point in Ca2+ homeostasis. The production of calcitriol is also increased by prolactin, a hormone which stimulates lactogenesis (the formation of breast milk), a process which requires large amounts of calcium. It is decreased by high levels of serum phosphate and by an increase in the production of the hormone FGF-23 by osteocyte cells in bone.

Aviva Systems Biology Calcitriol ELISA Kit (OKEH02542) is based on a competitive enzyme immunoassay technique. The microtiter well-plate in this kit has been pre-coated with an anti-Calcitriol antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated Calcitriol and incubated. The Calcitriol found in the sample or standards competes with the biotinylated Calcitriol for limited binding sites on the immobilized anti-Calcitriol antibody. Excess unbound biotinylated Calcitriol and sample or standard Calcitriol is washed from the plate. Avidin-HRP conjugate is added, incubated and washed. An enzymatic reaction is then produced through the addition of TMB substrate which is catalyzed by the immobilized HRP to generate a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration is measured by reading the absorbance at 450 nm which is quantitatively proportional to the amount of biotinylated Calcitriol captured in the well and inversely proportional to the amount of Calcitriol which was contained in the sample or standard.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol