How critical is the requirement for volumes of the sample to be 0.5 ml after diluting with Assay Buffer 13 0.5 ml at step 5 with Protein A ELISA kit, ADI-900-057?

Keeping the volume of sample to no more than 0.5ml is to ensure the entire Protein A in the sample would dissociate. The protocol for Protein A ELISA kit (ADI-900-057) has been optimized in these conditions. Boiling volumes greater than 0.5ml is not recommended. This would require optimization by the end user

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