The Human Vitamin D Binding Protein (DBP) ELISA, ECL Kit is a sensitive chemiluminescent sandwich enzyme immunoassay (ECL) for the in vitro quantitative measurement of DBP in human serum, plasma and other biological fluids.
Detection Range:0.47-30ng/ml
Sensitivity:< 0.13ng/ml
Precision:Intra-Assay: CV<10%Inter-Assay: CV<12%
Test Principle:The microtiter strips provided in this kit have been pre-coated with an antibody specific to DBP. A competitive inhibition reaction is launched between a fixed amount of biotin-labeled DBP and unlabeled DBP (in standards or samples) for limited binding sites on the pre-coated antibody. After incubation, the unbound conjugate is washed off. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. Finally, the mixture of substrate A and B is added to generate glow light emission kinetics and the chemiluminescence signal is read using a luminometer. The amount of bound HRP conjugate and the intensity of the emitted light are inversely proportional to the concentration of DBP in the sample or standard.
Kit Components:*158199A: Microtiter Plate, 1x96 wells, Pre-coated with antibody; ready to use*158199B: Standard, 2x1vial 158199C: Standard Diluent, 1x20ml*158199D: Detection Reagent A, 1x120ul *158199E: Detection Reagent B, 1x120ul 158199F: Assay Diluent A (2x), 1x6ml158199G: Assay Diluent B (2x),1x6ml158199H: Substrate A, 1x10ml158199K: Substrate B, 1x2ml158199L: Wash Buffer (30x), 1x20ml
Storage and Stability:Store *158199A, *158199B, *158199D and *158199E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Materials Required But Not Supplied:1. Luminometer capable of reading 96-well microplates with the following parameters: lag time 30.0secs; read time 1.0 sec/well. 2. Precision single or multi-channel pipettes and disposable tips
3. Eppendorf Tubes for diluting samples
4. Deionized or distilled water
5. Absorbent paper for blotting the microtiter plate
6. Container for Wash Solution
Assay Procedure Summary:1. Prepare all reagents, samples and standards
2. Add 100ul standard or sample to each well. Incubate 2 hours at 37°C
3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 100ul Substrate Solution. Incubate 10 minutes at 37°C
8. Read RLU immediately.
仕様
Size：96Tests
Source Antigen：Human
Specificity：This sandwich assay has high sensitivity and excellent specificity for detection of Vitamin D Binding Protein (DBP). No significant cross-reactivity or interference between Vitamin D Binding Protein (DBP) and analogues was observed.
