I am having an issue with an ELISA for mouse IgG: samples return lower values on the right side of the ELISA plate when compared to the left side of the ELISA plate. The difference is about 50%.
Although it is not our core business, I have done different ELISA's before with good results.
We have done some trouble shooting:
- We tested an in-house developed ELISA as well as 2 different commercial ELISA kits.
- The plate reader was excluded as the source of error.
- Temperature effects were excluded as the source of error.
Could this somehow be caused by the mouse IgG we want to measure, for example instability/denaturation of the protein in contact with air? Or am I doing something wrong?