ELISA is a common biological experiment. However, some problems may occur during the procedures. Some questions are answered below.

1. Why happened the phenomenon that the whole Micro ELISA Plate does not show blue color when adding TMB?

① Different reagents were mixed.

② The reagents were missed during the experiment.

③ Reagents were added in the wrong procedures.

④ Kits were stored improperly or have passed their expiry date.

⑤ TMB has been metamorphous.

 Wash Buffer was incorrectly prepared.

⑦ The pH of the Wash Buffer was too high or low.

2. Why happened the phenomenon that the whole Micro ELISA Plate show deep blue when adding TMB?

① The reagents were contaminated during the experiment.

② The experimental reaction time was too long.

③ Samples were improperly collected or contaminated.

④ Inadequate washing when adding Wash Buffer.

⑤ The Competition method principle Kit was performed according to assay procedures of the Sandwich method principle Kit.

3. How to calculate/ deal with the raw datas?

It is strongly recommended that fitting the curve of concentration(X-axis )and corrected OD values( Y-axis )  based on the four-parameter nonlinear fitting equation by using  many softwares like Origin(Preferred Choice), CurveExpert and so on.