Conjugation of Proteins with Horseradish Peroxidase (HRP)


The intracellular pathway of endocytosed ligands can be followed by density shift methods. When the ligand is conjugated to an enzyme like horseradish peroxidase (HRP), the organelle can be made denser by the addition of a substrate for this enzyme, diaminobenzidine (DAB). When this substrate is metabolized in the presence of hydrogen peroxide (H2O2), an insoluble aggregate is formed and the organelle will increase in density.

Keywords: endocytosis; cell fractionation; peroxidase; density shift; lysosome; conjugation


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Gjøen, Tor, and Berg, Trond(Jul 2003) Conjugation of Proteins with Horseradish Peroxidase (HRP). In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0003770]