VD3 ELISA kit (General species)

SKU: CEA920Ge

Size: 96Tests

# of Times Cited in literature: 2

Prepare Time: 1-3 days(please inquire for mutiple units)

Target Name: VD3

Target Full Name: Vitamin D3

Alternative Names: Cholecalciferol; Toxiferol; Activated 7-Dehydrocholesterol

Target Species: General species

Uniprot: -

Gene ID: -

Featured Series: CE kit

Featured Series Function: Detects small molecule

Specificity: Reactive with General species VD3 / Vitamin D3

Method: Colormetric

Detection principle: Competitive Inhibition

Detection range: 4.94-400ng/mL

Sensitivity: 2.14ng/mL

Assay Time: 2h

Sample Size: 50uL

Recommended/Predicted Sample Types: Serum, Plasma and other Biological Fluids

Assay Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%

Reproducibility test menthod: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vitamin D3 (VD3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vitamin D3 (VD3) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100

Storage: 4°C for 1 month/ -20°C for long-term(One year within shelf life)

Shelf-life: 12 months

Specificity: This assay has high sensitivity and excellent specificity for detection of Vitamin D3 (VD3). No significant cross-reactivity or interference between Vitamin D3 (VD3) and analogues was observed.

Stability: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Assay procedure summary: 1. Prepare all reagents, samples and standards; 2. Add 50µL standard or sample to each well. And then add 50µL prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C; 3. Aspirate and wash 3 times; 4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C; 5. Aspirate and wash 5 times; 6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C; 7. Add 50µL Stop Solution. Read at 450 nm immediately.

Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Vitamin D3 (VD3) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Vitamin D3 (VD3) and unlabeled Vitamin D3 (VD3) (Standards or samples) with the pre-coated antibody specific to Vitamin D3 (VD3). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Vitamin D3 (VD3) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Vitamin D3 (VD3) in the sample.

Research Area: Metabolic pathway;Reproductive science;Nutrition metabolism;Bone metabolism;

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