FAQ for ELISA kit operations:
1. Edge effect In the ELISA assay using a 96-well plate, an "edge effect" is often found, ie, the peripheral pores are darker than the central pores. Studies have shown that thermodynamic gradients in incubation may be the root cause. Polystyrene itself is a poor thermal conductor. In the laboratory routine ELISA, the plate is placed in a 37 ° C incubator from room temperature (usually around 25 ° C). When the plate is also warmed, it may be between the peripheral hole and the center hole. There is a thermodynamic gradient. Therefore, when a water bath is used or when the reaction solution is added to the pores of the plate, both the plate and the solution are heated to an incubation temperature (e.g., 37 ° C), the "edge effect" can be easily eliminated, and the repeatability of the measurement can be improved.
2. Loading? In the current ELISA commercial kit, there is bound to be a step of adding a sample using a micropipette. The key point of attention is: do not add too fast, avoid adding to the upper part of the hole wall, do not spill and create bubbles. Loading too fast, can not guarantee the accuracy and uniformity of micro-dosing. The non-coated area applied to the upper part of the pore wall is liable to cause non-specific adsorption. Spilling can contaminate adjacent holes. When bubbles appear, there is a difference in the interface of the reaction solution. Therefore, sometimes a specimen is tested positive with the same kit, and the next measurement is negative, which is often caused by the above-mentioned errors in loading and reagents.
Note:
1. When using the kit for the first time, the user should centrifuge the various reagent tubes for a few minutes to concentrate the reagents on the bottom of the tube.
2. Leave one well for each experiment as a blanking zero hole. This well is not filled with any reagents, only the final substrate solution and 2N H2SO4. Use this hole to adjust the OD value to zero when measuring.
3. To prevent the sample from evaporating, place the reaction plate in a closed box covered with a damp cloth and add a cover or a film to the plate.
4. Unused plate or reagent, please store at 2-8 °C. Standard, biotinylated antibody working solution, horseradish peroxidase labeled avidin working solution should be used according to the required amount. Do not reuse diluted standards, biotinylated antibody working solutions, or horseradish peroxidase-labeled avidin working solutions.
5. It is recommended to set the double hole measurement when testing the sample to ensure the accuracy of the test results.
(Note: This reagent is only for scientific research, please contact us!)
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