1,25-dihydroxyvitamin D3 ELISA Kit (Mouse) (OKCA00159)

• Product Number: OKCA00159
• Name: 1,25-dihydroxyvitamin D3 ELISA Kit (Mouse) (OKCA00159)
• Alias Symbols: Calcitriol, 1,25 DHVD3, 1,25-dihydroxyvitamin D3
• Description of Target: 1,25-Dihydroxyvitamin D3 is the physiologically active form of vitamin D. It is formed primarily in the kidney by enzymatic hydroxylation of 25-hydroxycholecalciferol (CALCIFEDIOL). Its production is stimulated by low blood calcium levels and parathyroid hormone. Calcitriol increases intestinal absorption of calcium and phosphorus, and in concert with parathyroid hormone increases bone resorption.
• Reconstitution and Storage: Store as indicated in product manual.
• Datasheets/Manuals: Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
• Specificity: This assay has high sensitivity and excellent specificity for detection of mouse
  DHVD 3. No significant cross-reactivity or interference between mouse DHVD 3
  and analogues was observed.
• Additional Information:

  CAS#: 32222-06-3

  Molecular Formula: C27H44O3

  Molecular Weight: 416.63646 g/mol

• Assay Info: Assay Methodology: Quantitative Competitive Immunoassay
• Sensitivity: < 250 fmoL/L
• Protein Name: 1,25-Dihydroxy vitamin D3
• Sample Type: Serum
• ELISA Kit Component:

  Component	Amount
  Anti-1,25-dihydroxyvitamin D3 Microplate	96 Well (12 x 8 Well Strips)
  1,25-dihydroxyvitamin D3 Standard	5 x 500 uL
  High Value Control	1 x 500 uL
  Middle Value Control	1 x 500 uL
  Sample Diluent	1 x 50 mL
  Anti-1,25-dihydroxyvitamin D3 Antibody HRP Conjugate	1 x 7 mL
  20X Wash Buffer	1 x 30 mL
  Substrate A	1 x 7 mL
  Substrate B	1 x 7 mL
  Stop Solution	1 x 7 mL

• ELISA Kit Reproducibility:

  	Mean Intra-Assay Precision %CV			Mean Inter-Assay Precision %CV
  Sample	1	2	3	1	2	3
  n	20	20	20	20	20	20
  Mean (fmol/L)	1996.76	2052.86	2097.57	2033.22	1934.69	2016.14
  SD	0.073	0.076	0.055	0.076	0.081	0.085
  CV%	5.647	6.018	4.397	5.974	6.197	6.669

• ELISA Kit Range: 1,000 fmol/L - 5,000 fmol/L
• ELISA Kit Duration: < 4 Hours
• ELISA Kit Principle: Aviva Systems Biology 1,25-dihydroxyvitamin D3 ELISA Kit (Mouse) (OKCA00159) is based on a competitive enzyme immuno assay technique. The microtiter well-plate in this kit has been pre-coated with Mouse 1,25-dihydroxyvitamin D3 and blocked. Standards or samples are added to the plate wells along with a horseradish peroxidase (HRP) conjugated antibody specific to 1,25-dihydroxyvitamin D3 and incubated. 1,25-dihydroxyvitamin D3 immobilized in the wells competes with the liquid phase 1,25-dihydroxyvitamin D3 contained in the samples or standards for binding with the HRP-Conjugated 1,25-dihydroxyvitamin D3 Antibody. Wells are washed removing unbound 1,25-dihydroxyvitamin D3, unbound HRP-Conjugated 1,25-dihydroxyvitamin D3 Antibody or 1,25-dihydroxyvitamin D3/HRP-Conjugated 1,25-dihydroxyvitamin D3 Antibody complex. HRP-Conjugated 1,25-dihydroxyvitamin D3 Antibody bound to 1,25-dihydroxyvitamin D3 immobilized in the well remains. An enzymatic reaction is 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 HRP-Conjugated 1,25-dihydroxyvitamin D3 Antibody captured in the well and inversely proportional to the amount of 1,25-dihydroxyvitamin D3 which was contained in the sample or standard.
• ELISA Kit Linearity:

  Sample Type 1
  Sample Type	Dilution	Average %	Range %
  Serum (n=4)	1:10	89%	83-96%
  Serum (n=4)	1:20	90%	85-95%
  Serum (n=4)	1:40	95%	90-99%
  Serum (n=4)	1:80	99%	96-104%

• ELISA Kit Detection Method: Colorimetric, OD 450 nm
• Gene Symbol: 1,25-dihydroxyvitamin D3
• Predicted Species Reactivity: Mouse
• Image 1: