As we approach the end of this month, there's only one week left. Our technical expert has conducted an experimental analysis and provided a detailed summary on the classic ELISA kit, focusing specifically on the blocking step.
Commonly used blocking agents include 0.05% to 0.5% BSA, 10% calf serum, or 1% gelatin. Skim milk powder is also widely used due to its affordability and effectiveness at higher concentrations (5% to 10%). Other options may include rare animal sera, casein, and more. While theoretical knowledge is important, practical testing is essential to determine what works best for your specific experiment. It’s always recommended to test different blocking agents to see which one yields the best results in your setup.
For coating solutions, common choices are phosphate buffer at pH 7.2, Tris-HCl buffer at pH 7–8, and carbonate buffer at pH 9.6. The purpose of blocking is to fill any remaining binding sites on the plate with unrelated proteins, preventing non-specific binding of interfering substances during later steps in the ELISA process. In some cases, the original coating might be blocked using a neutral buffer if it helps maintain the specificity of the target antigen.
Understanding these details can significantly improve the accuracy and reliability of your ELISA results. If you want to learn more about ELISA kits and related techniques, feel free to contact our company directly or leave a message on our website. We’re here to help and provide further guidance tailored to your needs.
Whether you're new to ELISA or looking to refine your technique, staying informed and experimenting with different methods will help you achieve better outcomes in your research.
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