Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
Principle of the Assay
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Caffeine. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Caffeine. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Caffeine in the samples is then determined by comparing the OD of the samples to the standard curve.
For use with: Serum, plasma, and other biological fluids.
For Research Use Only. Not for use in diagnostic procedures.
Target Information
| SYNONYMS | Caffeine |
Materials Supplied
| Kit Components | 96 Wells Quantity/Size |
|---|---|
| Pre-Coated Microplate | 48T: 6 strips x 8 wells; 96T: 12 strips x 8 wells |
| Standard (Lyophilized) | 48T: 1 vial; 96T: 2 vials |
| Biotinylated-Conjugate (100×) | 48T: 30 μL; 96T: 60 μL |
| Streptavidin-HRP (100×) | 48T: 60 μL; 96T: 120 μL |
| Standard/Sample Diluent Buffer | 48T: 10 mL; 96T: 20 mL |
| Biotinylated-Conjugate Diluent | 48T: 5 mL; 96T: 10 mL |
| HRP Diluent | 48T: 6 mL; 96T: 12 mL |
| Wash Buffer (25×) | 48T: 10 mL; 96T: 20 mL |
| TMB Substrate Solution | 48T: 6 mL; 96T: 10 mL |
| Stop Reagent | 48T: 3 mL; 96T: 6 mL |
| Plate Covers | 48T: 1 piece; 96T: 2 pieces |
Storage
| Storage | *If the kit is opened, Store the whole kit at 4°C. If the kit is not used up in 1 week. Store the Pre-Coated Microplate, Standard ,Biotinylated-Conjugate and Streptavidin-HRP at -20°C, the rest reagents at 4°C, please used up within 6 months. *If the kit is not opened, store the whole kit: 4°C(short time storage, valid for 6 months); -20°C (long-term storage, valid for 1 year). Avoid repeated freeze-thaw cycles |
Performance Characteristics
| REPEATABILITY | Intra-assay Precision (Precision within an assay)：CV%<8% Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays)：CV%<10% Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. |
| SENSITIVITY | The minimum detectable dose was 0.25ng/mL. |
| ASSAY RANGE | 0.47–30ng/mL |
| SPECIFICITY | This assay has high sensitivity and excellent specificity for detection of Caffeine. No significant cross-reactivity or interference between Caffeine and analogues was observed. |