Cell penetrating peptide for transduction of peptides and proteins into live cells
For in vitro use only!
Shipping: shipped on blue ice
Storage Conditions: store at -20 °C
Shelf Life: 12 months after date of delivery
Molecular Weight: 2247 Da confirmed by MALDI-MS.
Purity: > 95 % (HPLC)
Form: Synthetic peptide, water soluble powder, contains CF3COO- (trifluoro acetate) as counter ion.
Activity: 1 μl of stock solution is able to form a non-covalent complex with 1 μg of a protein of MW of 100 kDa. 10 to 20 μl of stock solution are necessary for complexation of 1 μg of a nucleoside triphosphate (MW approx. 0.5 kDa carrying 4 negative charges). For different MWs and/or different charges adjust amount of stock solution accordingly.
Penetratin is a cell penetrating peptide from the first generation, which is derived from Drosophila Antennapedia Homeodomain. It contains a nuclear localization sequence and facilitates internalization of cargo into living cells. Transport of the cargo requires in most cases the formation of a conjugate or fusion protein. Addition of a 10- to 20-fold excess of free penetratin to these constructs increases rate and efficiency of internalization. In some cases penetratin is able to form non-covalent complexes with the cargo. Penetratin shows only small cytotoxic effects on a number of cell lines (including HeLa, Jurkat, Swiss 3T3, NIH 3T3, NB-4, HT-1080, COS-7 and Leishmania tarentolae) and can used for internalization of proteins, nucleic acids as well as single nucleotides and nucleotide analogs. For cell survival the critical concentration of Penetratin in serum-free transduction medium is in the range of 20 μg/ml at which cell viability and cell membrane integrity are only marginally reduced (approx. 10-30 %).
Peptide provides 8 positive charges for complex formation. Due to ε-amino and guanido groups up to 11 trifluoro acetate residues may be present resulting in an apparent MW of about 3.1 kDa.
Dissolve 0.5 mg in 1.5 ml sterile and oxygen-free water according to the general manual. Use the solution immediately or aliquot and store at -20 °C. Avoid freeze / thaw cycles. Please note that the peptide may form S-oxide (Met) when stored in solution.
Perform calculation, complex formation and cargo transduction according to detailed protocols given in the general manual.
Jena Bioscience Publications using Penetratin:
Formation of non-covalent complexes with different cargos, transport into different cell lines, uptake efficiencies and cytotoxicity's are described in four publications:
Mussbach et al. (2011). Internalization of nucleoside phosphates into live cells by complex formation with different cell penetrating peptides and JBS-Nucleoducin. In: Langel U., Editor. Cell penetrating peptides Methods and Protocols. Methods in Molecular Biology, vol. 683, Humana Press, Springer, New York, Dordrecht, Heidelberg, London. pp. 375-389.
Mussbach et al. (2011). Transduction of peptides and proteins into live cells by cell penetrating peptides. J. Cell. Biochem. 112: 3824.
Keller et al. (2013). Relationships between cargo, cell penetrating peptides and cell type for uptake of non-covalent complexes into live cells. Pharmaceuticals 6: 184.
Keller et al. (2014). Transduction of proteins into Leishmania tarentolae by formation of non-covalent complexes with cell-penetrating peptides. J. Cell. Biochem. 115: 243.
Handbook of Cell-Penetrating Peptides, Second Edition, Ed. by Ü. Langel, CRC Taylor and Francis, Boca Raton, London, New York (2007).
Cell-Penetrating Peptides, Methods and Protocolls, Edited by Ülo Langel, Methods in Molecular Biology 683, Springer New York, Dodrecht, Heidelberg, London (2011).
Pharmaceuticals, Special Issue 'Cell penetrating Peptides' (2010-2013).
Morris et al. (2008) Cell penetrating peptides: from molecular mechanisms to therapeutics Bio.Cell 100:201.
Gros et al. (2006) A non-covalent peptide-based strategy for protein and peptide nucleic acid transduction. Biochim. Biophys. Acta 1758:384.
Lundberg et al. (2002) Positively charged DNA-binding proteins cause apparent cell membrane translocation. Biochem. Biophys. Res. Comm. 291:367.
Balayssac et al. (2006) Comparison of Penetratin and other homeodomain-derived cell-penetrating peptides: interaction in a membrane-mimicking environment and cellular uptake efficiency. Biochemistry 45:1408.
Albrizio et al. (2006) Driving force in the delivery of Penetratin conjugated G Protein fragment. J. Med. Chem. 50:1458.
Yesylevsky et al. (2009). Alternative mechanisms for the interaction of the cell-penetrating peptides penetratin and the TAT peptide with lipid bilayers. Biophysical J. 97: 40.
Ghibaudi et al. (2005). The interaction of the cell-penetrating peptide penetratin with heparin, heparansulfates and phospholipid vesicles investigated by ESR spectroscopy. J. Peptide Sci. 11: 401.
Ryves and Harwood (2006). Use of a penetratin-linked peptide in Dictostelium. Mol. Biotech-nology 33: 123.
Availability Restriction: Exclusively distributed in Japan by Greiner BioOne