Mouse histamine (HIS) ELISA enzyme-linked immunosorbent assay technology manufacturers also offer a wide range of related products, including ELISA kits, immunohistochemistry kits, cell culture media, antibodies, and standard solutions. These products are designed with high repeatability and reliability, ensuring accurate and consistent results. When purchasing an ELISA kit, you can expect professional, standardized, and efficient technical support to assist with your experiments.
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Here is a detailed step-by-step guide for performing the mouse histamine ELISA:
1. **Standard Dilution and Loading**: Prepare 10 standard wells on the ELISA plate. Add 100 μl of the standard solution into the first and second wells. Then add 50 μl of standard diluent into each of these two wells and mix thoroughly. Next, transfer 100 μl from the first and second wells into the third and fourth wells, respectively, and add 50 μl of diluent to each. Mix again. Discard 50 μl from the third and fourth wells, then add 50 μl of the standard solution into the fifth and sixth wells, followed by 50 μl of diluent. Repeat this process for the seventh, eighth, ninth, and tenth wells. After mixing, discard 50 μl from the ninth and tenth wells. The final concentrations in each well will be 12 μg/L, 8 μg/L, 4 μg/L, 2 μg/L, and 1 μg/L, respectively.
2. **Sample Addition**: Set up blank control wells (without sample or enzyme reagent), and add 40 μl of sample diluent to each test well. Then add 10 μl of the sample to be tested, resulting in a 5-fold dilution. Carefully pipette the sample into the bottom of each well, avoiding contact with the walls. Gently mix the contents.
3. **Incubation**: Seal the plate with an adhesive film and incubate at 37°C for 30 minutes.
4. **Washing Solution Preparation**: Dilute the concentrated washing solution 20 times with distilled water (for a 48T kit). Set aside for use.
5. **Washing**: Remove the sealing film, discard the liquid, and blot dry. Fill each well with washing solution, let it sit for 30 seconds, then discard. Repeat this process five times, and pat the plate dry.
6. **Enzyme Conjugate Addition**: Add 50 μl of enzyme-labeled reagent to each well, except for the blank control wells.
7. **Second Incubation**: Seal the plate again and incubate at 37°C for 30 minutes.
8. **Second Washing**: Follow the same washing procedure as step 5.
9. **Color Development**: Add 50 μl of color developer A, followed by 50 μl of color developer B. Gently mix the solution and avoid exposure to light. Incubate at 37°C for 15 minutes.
10. **Stop Reaction**: Add 50 μl of stop solution to each well to terminate the reaction. The color will change from blue to yellow.
11. **Absorbance Measurement**: Measure the OD values at 450 nm using a microplate reader. Ensure measurements are taken within 15 minutes after adding the stop solution.
By following these steps, you can accurately detect histamine levels in mouse samples. For more information or assistance, feel free to contact us!
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