At Which Stage Should Protein Concentration Be Determined After Extraction or After Reduction and Alkylation?
Protein concentration is typically determined immediately after protein extraction, prior to any subsequent procedures such as reduction and alkylation. This practice ensures accurate determination of the initial protein concentration for use in downstream experimental steps, thereby facilitating precise calculation and adjustment of the required reagent volumes. Following reduction and alkylation procedures, protein concentration is generally not reassessed, unless specifically required by the experimental design.
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