Clarify why the HRP-conjugate is added after the washing

2025-07-14

Question:" Clarify why the HRP-conjugate is added after the washing?"
 
Reply: The operational logic of ELISA follows a cycle of "binding - washing - rebinding - re - washing". Washing the plate aims to "purify" the reaction system and remove interferences.
As a signal marker, HRP must be added after plate washing to ensure that it only binds to the specifically bound target molecules. Eventually, it produces a real and quantifiable
signal through enzymatic reaction, which is a key step to ensure the sensitivity and specificity of ELISA detection.
 

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