1,25-dihydroxyvitamin D3,DVD/DHVD 3 ELISA Kit

Antigen: 1,25-dihydroxyvitamin D3,DVD/DHVD 3

Reaktivität: Human

Applikation: ELISA

Produktnummer: ABIN661411

Anwendungen

Protokoll: 1. Preparing reagents, samples and standards.
2. Preparing reagents, samples and standards, antibodies labeled with enzyme, reacting 60 minutes at 37 °C.
3. Plate washed five times, adding Chromogen solution A, B, reacting 10 minutes at 37 °C.
4. Add stop solution.
5. Measure the OD value within 10 minutes.
6. Calculation.

Aufbereitung der Reagenzien: Standard dilution: This test kit will be supplied with one original Standard reagent, please dilute it by yourself according to the instruction.
Standard No. 5: 120μl Original Standard + 120μl Standard diluent.
Standard No. 4: 120μl Standard No.5 + 120μl Standard diluent.
Standard No. 3: 120μl Standard No.4 + 120μl Standard diluent.
Standard No. 2: 120μl Standard No.3 + 120μl Standard diluent.
Standard No.1: 120μl Standard No.2 + 120μl Standard diluent.

Washing Method:
1. Manually washing method: Shake away the remain liquid in the enzyme plates, place some bibulous papers on the test-bed, and flap the plates on the upside down strongly. Inject at least 0.35ml after-dilution washing solution into the well, and marinate 1 to 2 minutes. Repeat this process according to your requirements.
2. Automatic washing method: If there is an automatic washing machine, it should only be used in the test when you are quite familiar with its function and performance.

Aufbereitung der Proben: Specimen requirements:
1. Don’t detect the samples with NaN3 which could inhibit HRP activity of the horseradish peroxidase.
2. Extract as soon as possible after Specimen collection. The extraction should conform with the relevant literature, and then carry out the experiment as soon as possible. If it is impossible to test immediately, preserve the specimen in -20 °C environment And repeated freezing and thawing is forbidden.

Ergebnisberechnung: [Suggestion:] If circumstances permit, the double-well assay is recommended for all the wells for more accurate results, accordingly the assay cost will increase. It is optional to assay the standard, and it is recommended to select the highest concentration, the lowest concentration and any one of the medium concentrations as reference.
1. Calculate the O.D values of all the wells with the standard and the wells with the samples to be assayed. Calculate average O.D values of all the wells with the standard and the wells with the sample to be assayed if the double-well assay is used.
2. Make the standard curve diagram with concentration of the standard from the low to the high and the left to the right as abscissa [X] axis and O.D values of the wells at 450nm as ordinate [Y] axis. In the [Logit-Log] linear regression model to map out a standard curve. The standard curve diagram can vary due to difference of laboratory conditions and other environmental factors at various regions. It is required to draw as many points as possible on the standard curve diagram, and other points are evenly distributed on both sides of the standard curve, thus ensuring more accurate assay results.
3. Find out the corresponding concentration range of each sample to be assayed on the standard curve diagram according to their O.D values. [Suggestion:] It is wise to use professional drawing software for analysis, find out the corresponding concentrations from the standard curve according to the O.D values of the samples to be assayed or calculate a regression equation of the standard curve according to the standard concentrations and the O.D values, and substitute the O.D value of the samples to be assayed in the equation to calculate the concentration.
4. Dilute the samples to the concentration range permitted by the kit for assay if the concentration of the sample is too high and beyond the range of the kit. And calculate the actual concentration by multiplying the result by the dilution folds.

Handhabung: Precautions:
1. Being taken out from the 2-8°C environment, the kit should be balanced 30 minutes in the ambient temperature then use. If the Coated plates of Enzyme haven’t been used up after opened, the remaining plates should be stored in Sealed bag.
2. For each step, add Sample with sample injector which should be calibrated frequently, in order to avoid unnecessary experimental tolerance.
3. The operation shall be carried out accordance to the instructions strictly. And test results must be based on the readings of the Enzyme reader.
4. In order to avoid cross-contamination, it is forbidden to re-use the suction head and seal plate membrane in your hands.
5. The idle agents shall be put up or covered. Do not use reagent with different batches. And use them before expired date.
6. The substrate B is light-sensitive. Prolonged exposure to light is forbidden.

Attention:
1. Certain technical risks may exist in the product as it is impossible to make more comprehensive identification and assay on raw materials under the existing technologies and conditions.
2. The final assay results are closely associated with operation levels of operators, laboratory conditions, ambient conditions, etc. Please prepare for sufficient samples for the sake of assay failure.
3. Do not expose the reagents to strong light or assay under the strong light in the storage and incubation procedures. Closely cap the bottles after use of the reagents to prevent evaporation or pollution.
4. The best assay results can be achieved in case of using all the reagents in the kit and strictly follow the instructions of the kit. All the reagents are associated and shall not be mixed with products of other manufacturers.

Benötigtes Material: 37°C incubator
Standard Enzyme reader
Precision pipettes and disposable pipette tips
distilled water
disposable tubes for sample dilution
absorbent paper

Lagerung: 1. The kit shall be stored at 2-8 °C, and the coated microwell plate shall be stored at a dry place.
2. The reagents shall be kept stable in the period of validity, and Substrate shall be colorless. The Substrate shall be changed in time in case of development or blueing.
Expiration: Twelve months (see label on the outer box for the specific date).

Beschränkungen: Nur für Forschungszwecke einsetzbar