Human Dopamine Receptor D2 (DRD2) Kit

This kit is only for in vitro research use and is not intended for clinical diagnosis

This kit is used for in vitro quantitative detection of human dopamine receptor D2 (DRD2) content in serum, plasma, tissue homogenates, and related liquid samples.

Validity period:6 months

Storage conditions:2-8℃

Kit composition:

Note:

1. (96T/48T) After opening the packaging, please check in a timely manner whether all items are complete.

2. The standard concentrations are: 800, 400, 200, 100, 50, and 25 pg/mL, respectively

3. After a large number of normal specimen tests, the normal concentration values of the specimens are within the detection range provided by the kit. During the experiment, 50 μ L of the sample can be directly taken and loaded. When some sample values exceed the standard concentration, the sample can be appropriately diluted with sample diluent before conducting the experiment.

Preparation before testing:

Please remove the reagent kit from the refrigerator 20 minutes in advance and allow it to equilibrate to room temperature.

2. The concentrated detergent taken out of the refrigerator may have crystallization, which is a normal phenomenon; Let it stand at room temperature, shake it gently until it dissolves, and then prepare the washing solution. 20ml of concentrated washing solution can be diluted with distilled water or deionized water to prepare 400ml of working concentration washing solution. The unused solution should be returned to 4 ℃

3.20 x dilution of washing buffer: Dilute distilled water at a ratio of 1:20, that is, 1 part of 20 x washing buffer is added to 19 parts of distilled water.

Human Dopamine Receptor D2 (DRD2) ELISA Kit

Experimental principle:

The reagent kit adopts a double antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA). Add the sample, standard, and HRP labeled detection antibody in sequence to the pre coated micropores with human dopamine receptor D2 (DRD2) capture antibodies, incubate and wash. Using substrate TMB for color development, TMB is converted to blue under the catalysis of peroxidase and to yellow under the action of acid. The depth of color is positively correlated with the human dopamine receptor D2 (DRD2) in the sample. Measure the absorbance (OD value) at a wavelength of 450nm using an enzyme-linked immunosorbent assay (ELISA) reader and calculate the sample concentration.

Self provided experimental equipment required for the experiment:

1. ELISA reader (450nm)

2. High precision sampler and nozzle: 0.5-10uL, 2-20uL, 20-200uL, 200-1000uL

3.37 ℃ Constant Temperature Chamber

4. Distilled water or deionized water

Sample processing and requirements:

1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or overnight at 4 ℃, then centrifuge at 1000 × g for 20 minutes to obtain the supernatant, or store the supernatant at -20 ℃ or -80 ℃, but avoid repeated freezing and thawing.

2. Plasma: Collect specimens using EDTA or heparin as anticoagulants, and centrifuge the specimens at 2-8 ℃ 1000 × g for 15 minutes within 30 minutes after collection. The supernatant can be collected for detection, or stored at -20 ℃ or -80 ℃, but repeated freezing and thawing should be avoided.

3. Tissue homogenate: Rinse the tissue with pre cooled PBS (0.01M, pH=7.4) to remove residual blood (lysed red blood cells in the homogenate can affect measurement results), weigh and cut the tissue into pieces. Mix the shredded tissue with the corresponding volume of PBS (usually in a weight to volume ratio of 1:9, for example, 1g of tissue sample corresponds to 9mL of PBS, the specific volume can be adjusted appropriately according to experimental needs, and records should be kept. Recommend adding protease inhibitors to PBS and grinding thoroughly on ice in a glass homogenizer. To further lyse tissue cells, the homogenate can be sonicated or subjected to repeated freeze-thaw cycles. Centrifuge the homogenate at 5000 × g for 5-10 minutes, and take the supernatant for detection.

4. Cell culture supernatant: Centrifuge at 1000 × g for 20 minutes, take the supernatant for detection, or store the supernatant at -20 ℃ or -80 ℃, but avoid repeated freezing and thawing.

5. Other biological specimens: Centrifuge at 1000 × g for 20 minutes and collect the supernatant for detection.

6. Sample appearance: The sample should be clear and transparent, and suspended solids should be removed by centrifugation.

7. Sample storage: If the sample is collected and tested within one week, it can be stored at 4 ℃. If it cannot be tested in a timely manner, it should be packaged according to a single use amount and frozen at -20 ℃ (tested within one month) or -80 ℃ (tested within six months) to avoid repeated freezing and thawing. Hemolysis of the sample will affect the test results, so hemolytic samples should not be tested for this purpose.

Notes:

1. Strictly follow the prescribed time and temperature for incubation to ensure accurate results. All reagents must reach room temperature of 20-25 ℃ before use. Immediately refrigerate and store the reagents after use.

Incorrect board washing can lead to inaccurate results. Ensure to absorb as much liquid as possible from the well before adding the substrate. Do not let the micropores dry out during the incubation process.

3. Eliminate residual liquid and fingerprints at the bottom of the board, otherwise it will affect the OD value.

4. The substrate color solution should be colorless or very light in color, and the substrate solution that has turned blue cannot be used.

5. Avoid cross contamination between reagents and specimens to prevent erroneous results.

6. Avoid direct exposure to strong light during storage and incubation.

7. After balancing to room temperature, open the sealing bag to prevent water droplets from condensing on the cold Flat noodles.

8. Any reaction reagents should not come into contact with bleach solvents or the strong gases emitted by bleach solvents. Any bleaching component will destroy the biological activity of the reactants in the reagent kit.

9. Expired products cannot be used.

10. If there is a possibility of spreading the disease, all samples should be managed and processed according to the prescribed procedures and testing devices.

Operation steps:

1. Take out the required Flat noodles from the aluminum foil bag after 60 min of room temperature balance, and seal the remaining Flat noodles with a self sealing bag and put it back at 4 ℃.

2. Set up standard wells and sample wells, and add 50 μ L of standard samples of different concentrations to each standard well.

3. Add 50 μ L of the test sample to the sample well a; Blank holes are not added.

4. Except for blank wells, add 100 μ L of horseradish peroxidase (HRP) labeled detection antibody to each well of the standard and sample wells, seal the reaction well with a sealing plate membrane, and incubate at 37 ℃ in a water bath or constant temperature box for 60 minutes.

5. Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (350 μ L), let it stand for 1 minute, shake off the washing solution, pat dry on absorbent paper, repeat washing the board 5 times (or use a washing machine to wash the board).

6. Add 50 μ L of substrate A and B to each well, and incubate at 37 ℃ in the dark for 15 minutes.

7. Add 50 μ L of termination solution to each well and measure the OD value of each well at a wavelength of 450nm within 15 minutes.

Experimental result calculation:

Draw standard curve: In an Excel worksheet, use the standard concentration as the horizontal axis and the corresponding OD value as the vertical axis to draw a linear regression curve of the standard. Calculate the concentration values of each sample according to the curve equation.

(This image is for reference only)

Human Dopamine Receptor D2 (DRD2) ELISA Kit

Performance:

1. Detection range: 25 pg/mL -800 pg/mL.

2. Sensitivity: The detection concentration is less than 1.0 pg/mL.

3. Specificity: Does not cross react with other soluble structural analogues.

4. Repeatability: The intra plate coefficient of variation is less than 10%, and the inter plate coefficient of variation is less than 15%.

Technical tip:

When mixing or redissolving protein solutions, try to avoid foaming as much as possible.

To avoid cross infection, it is necessary to replace the nozzle when configuring different concentration standards, loading samples, and adding different reagents. Additionally, please use different pipettes for different reagents.

3. During each incubation, please use the sealing glue correctly to ensure the accuracy of the results.

4. The mixed chromogenic substrate should be colorless before being loaded onto the plate. Please store it away from light; If the microplate changes from being searched to different depths of blue.

5. The order of terminating the liquid on the plate should be consistent with the order of the color developing substrate on the plate; After adding the termination solution, the color inside the hole changed from blue to yellow; If there is green inside the hole, it indicates that the liquid inside the hole is not mixed evenly; Please mix thoroughly.

Explanation:

Due to the current conditions and scientific and technological level, it is not yet possible to comprehensively identify and analyze all raw materials provided by all suppliers, which may pose certain quality and technical risks to this product.

2. The experimental results are closely related to the effectiveness of the reagents, the relevant operations of the experimenter, and the experimental environment at that time. Please be sure to prepare sufficient specimen backups.

3. There may be slight differences in the detection limit, sensitivity, and color development time of the same product from different batches. Please refer to the instructions in the reagent kit for experimental operations. The electronic version of the instructions on the website is for reference only.

4. Only by using all the reagents in this kit can the detection effect be guaranteed, and products from other manufacturers cannot be mixed. Only by strictly following the experimental instructions of this kit can the test results be obtained.