The ELISA method is a benchmark for quantitation of pathological antigens and there are indeed many variations to this method. ELISAs are adaptable to high-throughput screening because results are rapid, consistent and relatively easy to analyze. The best results have been obtained with the sandwich format, utilizing highly purified, prematched capture and detector antibodies. The resulting signal provides data which is very sensitive and highly specific.
This section is intended to recommend some techinical tips for the proper usage of ELISA kit.
1. Bring all reagents and samples to room temperature before use.
2. Samples should be thawed completely and mixed well prior to analysis. Avoid repeated freeze-thaw cycles of frozen samples.
3.A standard curve should be generated for each set of sample assayed. DO NOT USE the standard curves from other plates or other days.
4.Use a new disposable reagent reservoir and new disposable pipette tips for each transfer to avoid cross-contamination.
5.Read the absorbance of each well within 20 minutes after adding the stop solution.