Delivery of proteins to the cytosol of living cells is a promising research tool. Delivery of antibodies in particular bears exciting applications such as in vivo tracking of proteins at endogenous expression levels or interference with cellular processes. In spite of the large number of methods published for protein delivery, successful applications so far are rare. A possible explanation for this is a vast overestimation of the delivery efficiency due to the use of inappropriate detection methods and/or unsuitable positive controls for cytosolic delivery. Therefore, we provide strategies for unequivocally detecting cytoplasmic protein delivery and quantifying protein transformation efficiency. Finally, we present a protocol for efficient protein delivery to the cytosol validated using these methods.
Antibody Profection Protein transfection Protein delivery Cell-penetrating peptide (CPP) Protein transduction domain (PTD) Electroporation Yumab
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We are grateful to André Frenzel and Thomas Schirrmann for helpful discussions and the EU consortium “Affinomics” for financial support.
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