Current Position:Home>>Hot>>Immunology Related Reagents>>Other Reagents>> Biotin Quantitative Determination Kit
Catalog number:E-IR-R501
Application: ELISA
Storage: 2~8°C
Lead Time: Order now, ship in 3 daysWelcome to order from local distributors.
Add to cart Compare Bulk request ManualThis kit uses Competitive-method. The microplate provided in this kit has been pre-coated with Biotin. During the reaction, Biotin in the sample or standard competes with a fixed amount of Biotin on the solid phase supporter for sites on the Avidin-HRP specific to Biotin. Excess Avidin-HRP and unbound sample or standard are washed from the plate. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change can be measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Biotin in samples can be calculated by comparing the OD of the samples with the standard curve.
Calculation of results
Average the duplicate readings for each standard and samples. Plot a four-parameter logistic curve on log-log graph paper, with standard concentration on the x-axis and OD values on the y-axis.
If samples have been diluted, the concentration calculated from the standard curve must be multiplied by the dilution factor. If the OD of the sample surpasses the lower limit of the standard curve, you should re-test it after appropriate dilution. The actual concentration is the calculated concentration multiplied dilution factor.
SUMMARY
1. Add 50 μL standard or sample to each well.
2. Immediately add 50 μL Avidin-HRP to each well.3. Incubate for 30 min at 37℃
4. Aspirate and wash 5 times
5. Add 90 μL Substrate Reagent. Incubate 15 min at 37℃
6. Add 50 μL Stop Solution. Read at 450 nm immediately.
7. Calculation of results
Kit components & Storage
The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions since the kit is received.
Item |
Specifications |
Storage |
Micro ELISA Plate (Dismountable) |
8×12 or 8×6 * |
-20℃, 12 months |
Reference Standard |
2/1 vial |
|
Concentrated Avidin-HRP |
1 vial, 120 μL/60 μL * |
4℃(shading light), 12 months |
Reference Standard & Sample Diluent |
1 vial, 20 mL/12 mL * |
4℃, 12 months |
HRP Conjugate Diluent |
1 vial, 14 mL |
|
Concentrated Wash Buffer (25×) |
1 vial, 30 mL |
|
Substrate Reagent |
1 vial, 10 mL |
4℃(shading light) |
Stop Solution |
1 vial, 10 mL |
4℃ |
Plate Sealer |
5/3pieces * |
|
Product Description |
1 copy |
|
Certificate of Analysis |
1 copy |
Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use in measuring instead of directly pouring.
Other supplies required
Microplate reader with 450nm wavelength filter
High-precision transferpettor, EP tubes and disposable pipette tips
37°C Incubator, Deionized or distilled water
Absorbent paper