The attachment of a reporter group (label) is often required for the detection and biochemical/cellular characterization of proteins and its binding partners.
Different labeling techniques & types of label such as fluorophores, biotin and luminescent dyes are available for non-radioactive protein labeling, both random and site-directed (N- or C-terminal).
Choose the appropriate labeling strategy (labeling technique & type of label) that fits your final application based on your initial protein source (Tab. 1).
Table 1: Protein Labeling Selection Matrix
|Type of Label|
|Protein Source||Purified protein||Amine Labeling||Amine Labeling||Amine Labeling → Click Chemistry||random||Labeling Position|
|Thiol Labeling||Thiol Labeling → Click Chemistry|
|Proliferating cells (Nascent proteins)||Amino acid-based Metabolic Labeling|
|Puromycin-based Metabolic Labeling||site directed|
|ATP dependent Lysine-based Labeling||ATP dependent Lysine-based Labeling|
|Protein encoding DNA sequence (Cloning & cell-free expression required)||dC-Puromycin-based Co-translational Labeling||dC-Puromycin-based Co-translational Labeling|