2. Reagents preparation: Please finish preparing reagent 10 minutes before using. Centrifugation is required for all reagent components when using for the first time so as to concentrate them to the bottom of the tube. Please use precise measuring tools (pipette, graduated cylinder etc.) for reagent preparation, because the actual quantity provided for each reagent (eg: Wash Buffer,Sample Diluent) is more than what the label says. Do NEVER take the labeled quantity for granted and add solution directly into the vials for reagent preparation. All experimental apparatus should be clean and you’d better refer to the insert for details of reagent preparation.
3. ELISA plate: It’s OK that the aluminum foil bags is leaking since it won’t affect the quality of the plate. The strips are movable and it’s recommended to separate those wells needed for assay and store the rest at 2-8 ℃ in dark without germ contamination. Do NEVER expose the whole plate when only some of them are needed.
4. Samples or reagents adding : It is recommended that the whole time for sample adding should be controlled in 5 minutes and added volume for each well should be the same to keep reaction consistency.
5. Incubation: To avoid sample evaporation and contamination, proper adhesion of plate sealers during incubation steps is necessary. Be careful and do not spill the liquid when moving the plate. Since temperature stability is very important, please carefully monitor incubating temperature and keep it at 37℃. but avoid opening the door of incubator too many times during incubation.
6. Washing: Please ensure the same wash buffer volume per well and you’d better use multi-channel pipette. If you use the ELISA Washer, please make sure it’s clean enough without contamination. No rinse but gently wash. Throw plates vertically and wash it thoroughly to reduce the edge effect. For detailed procedure, please refer to the manual insert.
7. Reading: Please read the plate in 5 minutes after adding the stop solution to avoid generation of brown precipitate that may affect the result. Make sure that the microplate reader is set correctly and the filter is precisely calibrated.
8. Do not make mix use of reagents within different lot#. Also, cross-use of caps matched to different reagents is also forbidden. Do NEVER replace components of our kit with other’s.