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Home » Products and Applications » Delivery Products

Endo-Porter

Endo-Porter simple delivery reagent
  1. What is it and how does it work?
    2. Endo-Porter is a novel peptide* explicitly designed to do what all existing peptides and lipid-based delivery reagents can not: deliver substances into the cytosol of cells by an endocytosis-mediated process that avoids damaging the plasma membrane of the cell. This mechanism prevents the loss of vital cell contents with its associated toxicity. Endo-Porter requires no interaction with the cargo to be delivered and allows the cargo that is endocytosed along with Endo-Porter to exit the endosome and enter the cytosol. Detailed studies with a FITC-anti-actin antibody fragment (Sigma F-3046) and fluorescent high-molecular weight dextrans suggest Endo-Porter can deliver high concentrations of cargo exceeding 70kD into both adherent and non-adherent cells. Unlike the many delivery reagents that claim effectiveness in the presence of serum, Endo-Porter actually works with serum concentrations up to the 10% standard. Endo-Porter delivers substances such as peptides, proteins, antibodies and is excellent for delivering Morpholino antisense oligos as well! We do not recommend using Endo-Porter to deliver anionic nucleic acids, as delivery efficacy is poor. We think complexation of high-anionic-density compounds with the polycationic Endo-Porter interferes with the activity of Endo-Porter.

    * Patented: US 7,084,248 B2

    Figure: Endo-Porter delivery mechanism and image of Endo-Porter delivery of 70 kD dextran tagged with Texas Red

    View or download an animation of Endo-Porter's mechanism of action by selecting one of the following two choices:

    Windows Media Apple QuickTime

    Animation by Pathworks of Corvallis, OR.
  2. What are the advantages of Endo-Porter over other delivery reagents?
    3. Most other delivery reagents require pre-incubation or optimized reagent-to-cargo ratio in order to achieve delivery, but not Endo-Porter. You can add as much or as little cargo as desired and use the same amount of Endo-Porter EVERY time. Endo-Porter lasts indefinitely and can be stored at room temperature or below.
  3. The protocol is so simple it fits on the box!
    4. Our recommended delivery procedure:

    1. Replace spent culture medium with fresh medium (preferably 10% serum).
    2. Add the desired cargo and swirl to mix.
    3. Add 6 µL of Endo-Porter for every 1 mL of culture medium & cargo and immediately swirl to mix.

    In most cases, long-term (72 hour) and short-term (24 hour) delivery is optimal at 6 µM Endo-Porter (6 µL/mL); however, cell lines vary in their response to Endo-Porter and it is prudent to start experiments with a new cell line by doing a ranging study for efficacy and toxicity at 2, 4, and 6 µL/mL Endo-Porter. Check the cells at 72 hours for signs of toxicity, then use the highest concentration that the cells tolerate for subsequent delivery procedures. Unlike most delivery solutions, 10% serum concentrations yield better results than reduced serum levels. Additional cargo and Endo-Porter can be added with each media replacement if desired. For delivery of Morpholino oligos, we suggest you start with 10 µM oligo final concentration in the culture medium then adjust concentrations as needed to achieve effective and specific gene knockdown.

    Endo-Porter protocol (PDF)
  4. What do I get and how much does it cost?
    5. For $200 you will receive 1.0 mL of 1.0 mM Endo-Porter in a 100% DMSO solution. This is enough to deliver up to 160 mL of cells with Endo-Porter. Endo-Porter can be ordered online today!

    For additional questions or assistance from our Ph.D.-level customer support staff use our online live help, submit a question, or call (541) 929-7840 ext. 1.

  5. Endo-Porter references
    6.
    A description of the development of Endo-Porter has been published:

    Summerton JE. Endo-Porter: A Novel Reagent for Safe, Effective Delivery of Substances into Cells. Ann N Y Acad Sci. 2005 Nov;1058:62-75

    Long J, Wang Y, Wang W, Chang BH, Danesh FR. Identification of MicroRNA-93 as a novel regulator of vascular endothelial growth factor (VEGF) In hyperglycemic conditions. J Biol Chem. 2010 May 25. [Epub ahead of print]
    Abstract mentions peptide-conjugated Morpholino oligomers, but mat'l and methods describes Endo-Porter.

    Lahat G, Zhu QS, Huang KL, Wang S, Bolshakov S, Liu J, Torres K, Langley RR, Lazar AJ, Hung MC, Lev D. Vimentin is a novel anti-cancer therapeutic target; insights from in vitro and in vivo mice xenograft studies. PLoS One. 2010 Apr 16;5(4):e10105.

    Matluk N, Rochiera JA, Karaczyn A, Adams TL, Verdi JM. A role for NRAGE in NF-kappaB activation through the non-canonical BMP pathway. BMC Biol. 2010 Jan 25;8(1):7. [Epub ahead of print]

    Pillai VC, Yesudas R, Shaik IH, Thekkumkara TJ, Bickel U, Srivenugopal KS, Mehvar R. Delivery of NADPH-Cytochrome P450 Reductase Antisense Oligos Using Avidin-Biotin Approach. Bioconjug Chem. 2010 Jan 11. [Epub ahead of print]

    Ikeuchi N, Futami J, Hosoi A, Noji S, Kurachi M, Ueha S, Fujii SI, Yamada H, Matsushima K, Moriyasu F, Kakimi K. Efficient cross-presentation of soluble exogenous antigens introduced into dendritic cells using a weak-based amphiphilic peptide. Biochem Biophys Res Commun. 2010 Jan 11. [Epub ahead of print]

    Fercher A, Ponomarev GV, Yashunski D, Papkovsky D. Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes. Anal Bioanal Chem. 2010 Jan 10. [Epub ahead of print]

    Spigoni G, Gedressi C, Mallamaci A. Regulation of Emx2 expression by antisense transcripts in murine cortico-cerebral precursors. PLoS One. 2010 Jan 11;5(1):e8658.
    Morpholinos were delivered to NIH/3T3 cells.

    Figeac F, Uzan B, Faro M, Chelali N, Portha B, Movassat J. Neonatal growth and regeneration of {beta} cells are regulated by the Wnt/{beta}-catenin signaling in normal and diabetic rats. Am J Physiol Endocrinol Metab. 2009 Nov 17. [Epub ahead of print]

    Yew KH, Carsten B, Harrison C. Scavenger receptor A1 is required for sensing HCMV by endosomal TLR-3/-9 in monocytic THP-1 cells. Mol Immunol. 2009 Nov 13. [Epub ahead of print]

    Nossa CW, Jain P, Tamilselvam B, Gupta VR, Chen LF, Schreiber V, Desnoyers S, Blanke SR. Activation of the abundant nuclear factor poly(ADP-ribose) polymerase-1 by Helicobacter pylori. Proc Natl Acad Sci U S A. 2009 Nov 24;106(47):19998-20003. Epub 2009 Nov 6.

    Chen AY, Zhang EY, Guan W, Cheng F, Kleiboeker S, Yankee TM, Qiu J. The small 11kDa non-structural protein of human parvovirus B19 plays a key role in inducing apoptosis during B19 virus infection of primary erythroid progenitor cells. Blood. 2009 Oct 27. [Epub ahead of print]

    Maxhimer JB, Soto-Pantoja DR, Ridnour LA, Shih HB, DeGraff WG, Tsokos M, Wink DA, Isenberg JS, Roberts DD. Radioprotection in Normal Tissue and Delayed Tumor Growth by Blockade of CD47 Signaling. Sci Transl Med. 2009;1(3):3ra7

    Pérez B, Rincón A, Jorge-Finnigan A, Richard E, Merinero B, Ugarte M, Desviat LR. Pseudoexon exclusion by antisense therapy in methylmalonic aciduria (MMAuria). Hum Mutat. 2009 Sep 2. [Epub ahead of print]

    Okamoto M, Van Stry M, Chung L, Koyanagi M, Sun X, Suzuki Y, Ohara O, Kitamura H, Hijikata A, Kubo M, Bix M. Mina, an Il4 repressor, controls T helper type 2 bias. Nat Immunol. 2009 Aug;10(8):872-9. Epub 2009 Jun 28.
    Primary C57BL/6 CD4+ T cells

    Rodríguez-Pascau L, Coll MJ, Vilageliu L, Grinberg D. Antisense oligonucleotide treatment for a pseudoexon-generating mutation in the NPC1 gene causing Niemann-Pick type C disease. Hum Mutat. 2009 Aug 30. [Epub ahead of print]

    Lin Z, Murtaza I, Wang K, Jiao J, Gao J, Li PF. miR-23a functions downstream of NFATc3 to regulate cardiac hypertrophy. Proc Natl Acad Sci U S A. 2009 Jul 2. [Epub ahead of print]

    Liu B, Paranjpe S, Bowen WC, Bell AW, Luo JH, Yu YP, Mars WM, Michalopoulos GK. Investigation of the Role of Glypican 3 in Liver Regeneration and Hepatocyte Proliferation. Am J Pathol. 2009 Jul 2. [Epub ahead of print]

    Kamel S, Kruger C, Salbaum JM, Kappen C. Morpholino-mediated knockdown in primary chondrocytes implicates Hoxc8 in regulation of cell cycle progression. Bone. 2008 Nov 21. [Epub ahead of print]

    Nikopoulos GN, Martins JF, Adams TL, Karaczyn A, Adams D, Vary C, Verdi JM. NRAGE: A Potential Rheostat During Branching Morphogenesis. Mech Dev. 2009 May-Jun;126(5-6):337-49. Epub 2009 Mar 4.

    Aouadi M, Tesz GJ, Nicoloro SM, Wang M, Chouinard M, Soto E, Ostroff GR, Czech MP. Orally delivered siRNA targeting macrophage Map4k4 suppresses systemic inflammation. Nature. 2009 Apr 30;458(7242):1180-4.

    Wang X, Gocek E, Liu CG, Studzinski GP. MicroRNAs181 regulate the expression of p27Kip1 in human myeloid leukemia cells induced to differentiate by 1,25-dihydroxyvitamin D3. Cell Cycle. 2009 Mar 1;8(5):736-41. Epub 2009 Mar 18.
    MicroRNA precursors (pre-miRNA) delivered into HL60 and U937 cells.

    Uzan B, Figeac F, Portha B, Movassat J. Mechanisms of KGF Mediated Signaling in Pancreatic Duct Cell Proliferation and Differentiation. PLoS ONE. 2009;4(3):e4734. Epub 2009 Mar 6.

    Pros E, Fernández-Rodríguez J, Canet B, Benito L, Sánchez A, Benavides A, Ramos FJ, López-Ariztegui MA, Capellá G, Blanco I, Serra E, Lázaro C. Antisense therapeutics for neurofibromatosis type 1 caused by deep intronic mutations. Hum Mutat. 2009 Feb 24;30(3):454-462. [Epub ahead of print]

    O'Hagan KA, Cocchiglia S, Zhdanov AV, Tambawala MM, Cummins EP, Monfared M, Agbor TA, Garvey JF, Papkovsky DB, Taylor CT, Allan BB. PGC-1alpha is coupled to HIF-1alpha-dependent gene expression by increasing mitochondrial oxygen consumption in skeletal muscle cells. Proc Natl Acad Sci U S A. 2009 Feb 17;106(7):2188-93. Epub 2009 Jan 28.

    Göoz P, Göoz M, Baldys A, Hoffman S. ADAM-17 regulates endothelial cell morphology, proliferation, and in vitro angiogenesis. Biochem Biophys Res Commun. 2009 Feb 27;380(1):33-8. Epub 2009 Jan 14.

    Lynch VJ, Tanzer A, Wang Y, Leung FC, Gellersen B, Emera D, Wagner GP. Adaptive changes in the transcription factor HoxA-11 are essential for the evolution of pregnancy in mammals. Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):14928-33. Epub 2008 Sep 22.

    Davis RL, Homer VM, George PM, Brennan SO. A deep intronic mutation in FGB creates a consensus exonic splicing enhancer motif that results in afibrinogenemia caused by aberrant mRNA splicing, which can be corrected in vitro with antisense oligonucleotide treatment. Hum Mutat. 2008 Oct 13. [Epub ahead of print]

    Sun CS, Wu KT, Lee HH, Uen YH, Tian YF, Tzeng CC, Wang AH, Cheng CJ, Tsai SL. Anti-sense morpholino oligonucleotide assay shows critical involvement for NF-kappaB activation in the production of Wnt-1 protein by HepG2 cells: oncology implications. J Biomed Sci. 2008 Sep;15(5):633-43. Epub 2008 May 7.

    Staszkiewicz J, Gimble J, Manuel JA, Gawronska-Kozak B. IFATS Series: Stem Cell Antigen-1 Positive Ear Mesenchymal Stem Cells (EMSC) Display Enhanced Adipogenic Potential. Stem Cells. 2008 Jul 3. [Epub ahead of print]

    Chiu SL, Chen CM, Cline HT. Insulin receptor signaling regulates synapse number, dendritic plasticity, and circuit function in vivo. Neuron. 2008 Jun 12;58(5):708-19.

    Kahler RA, Yingst SM, Hoeppner LH, Jensen ED, Krawczak D, Oxford JT, Westendorf JJ. Collagen 11a1 is indirectly activated by lymphocyte enhancer-binding factor 1 (Lef1) and negatively regulates osteoblast maturation. Matrix Biol. 2008 May;27(4):330-8. Epub 2008 Jan 16.

    Isenberg JS, Romeo MJ, Maxhimer JB, Smedley J, Frazier WA, Roberts DD. Gene Silencing of CD47 and Antibody Ligation of Thrombospondin-1 Enhance Ischemic Tissue Survival in a Porcine Model: Implications for Human Disease. Annals of Surgery. 2008 May;247(5):860-868.

    Nikopoulos G, Adams T, Adams D, Oxburgh L, Prudovsky I, Verdi J. The use of Endo-Porter to deliver morpholinos in kidney organ culture. Biotechniques. 2008 Apr;44(4):547-9.

    Coate TM, Wirz JA, Copenhaver PF. Reverse Signaling via a Glycosyl-Phosphatidylinositol-Linked Ephrin Prevents Midline Crossing by Migratory Neurons during Embryonic Development in Manduca. J Neurosci. 2008 Apr 9;28(15):3846-60.

    Chiang PC, Hsu JL, Yeh TC, Pan SL, Guh JH. Elucidation of susceptible factors to endoplasmic reticulum stress-mediated anticancer activity in human hepatocellular carcinoma. Naunyn Schmiedebergs Arch Pharmacol. 2008 Apr;377(2):167-77. Epub 2008 Jan 29.

    Ptaszynska MM, Pendrak ML, Bandle RW, Stracke ML, Roberts DD. Positive Feedback between Vascular Endothelial Growth Factor-A and Autotaxin in Ovarian Cancer Cells. Mol Cancer Res. 2008 Mar;6(3):352-63.

    Yarwood SJ, Borland G, Sands WA, Palmer TM. Identification of CCAAT/Enhancer-binding Proteins as Exchange Protein Activated by cAMP-activated Transcription Factors That Mediate the Induction of the SOCS-3 Gene. J Biol Chem. 2008 Mar 14;283(11):6843-53. Epub 2008 Jan 14

    Islam KN, Mendelson CR. Glucocorticoid/GR Inhibition of Surfactant Protein-A (SP-A) Gene Expression in Lung Type II Cells is Mediated by Repressive Changes in Histone Modification at the SP-A Promoter. Mol Endocrinol. 2008 Mar;22(3):585-96. Epub 2007 Dec 13

    Inoue Y, Kurihara R, Tsuchida A, Hasegawa M, Nagashima T, Mori T, Niidome T, Katayama Y, Okitsu O. Efficient delivery of siRNA using dendritic poly(L-lysine) for loss-of-function analysis. J Control Release. 2008 Feb 18;126(1):59-66. Epub 2007 Nov 6.

    Laguinge LM, Samara RN, Wang W, El-Deiry WS, Corner G, Augenlicht L, Mishra L, Jessup JM. DR5 receptor mediates anoikis in human colorectal carcinoma cell lines. Cancer Res. 2008 Feb 1;68(3):909-17.

    Place RF, Li LC, Pookot D, Noonan EJ, Dahiya R. MicroRNA-373 induces expression of genes with complementary promoter sequences. Proc Natl Acad Sci U S A. 2008 Jan 28; [Epub ahead of print]

    Zhdanov AV, Ward MW, Prehn JH, Papkovsky DB. Dynamics of intracellular oxygen in PC12 cells upon stimulation of neurotransmission. J Biol Chem. 2007 Dec 17; [Epub ahead of print]

    Ridnour LA, Windhausen AN, Isenberg JS, Yeung N, Thomas DD, Vitek MP, Roberts DD, Wink DA. Nitric oxide regulates matrix metalloproteinase-9 activity by guanylyl-cyclase-dependent and -independent pathways. Proc Natl Acad Sci U S A. 2007 Oct 23;104(43):16898-903. Epub 2007 Oct 17.

    Wang H, Chen C, Song X, Chen J, Zhen Y, Sun K, Hui R. Mef2c is an essential regulatory element required for unique expression of the cardiac-specific CARK gene. J Cell Mol Med. 2007 Nov 16; [Epub ahead of print]

    Ugarte M, Aguado C, Desviat LR, Sanchez-Alcudia R, Rincon A, Perez B. Propionic and Methylmalonic Acidemia: Antisense Therapeutics for Intronic Variations Causing Aberrantly Spliced Messenger RNA. Am J Hum Genet. 2007 Oct 26;81(6) [Epub ahead of print]

    Mandayam S, Huang R, Tarnawski AS, Chiou SK. Roles of survivin isoforms in the chemopreventive actions of NSAIDS on colon cancer cells. Apoptosis. 2007 Jun;12(6):1109-16. Epub 2006 Dec 8.

    Kikuno N, Shiina H, Urakami S, Kawamoto K, Hirata H, Tanaka Y, Place RF, Pookot D, Majid S, Igawa M, Dahiya R. Knockdown of astrocyte-elevated gene-1 inhibits prostate cancer progression through upregulation of FOXO3a activity. Oncogene. 2007 Jun 11; [Epub ahead of print]

    Ammons MC, Siemsen DW, Nelson-Overton LK, Quinn MT, Gauss KA. Binding of Pleomorphic Adenoma Gene-like 2 to the Tumor Necrosis Factor (TNF)-{alpha}-responsive Region of the NCF2 Promoter Regulates p67phox Expression and NADPH Oxidase Activity. J Biol Chem. 2007 Jun 15;282(24):17941-52. Epub 2007 Apr 26.

    Challa P, Soto Leon FA, Talbot L, Liton PB, Luna C, Epstein DL, MaddalacR and Gonzalez P. Novel Anti-Sense Oligonucleotide Delivery to Primary Lens Epithelial and Trabecular Meshwork Cell Cultures. Invest Ophthalmol Vis Sci 2007;48: E-Abstract 4242.

    Choe KP, Stidham J, Evans DH. Progress towards a method of targeted protein knockdown in Fundulus heteroclitus. The Bulletin, MDI Biological Laboratory V. 45, 2006.

    Lu PH, Chueh SC, Kung FL, Pan SL, Shen YC, Guh JH. Ilimaquinone, a marine sponge metabolite, displays anticancer activity via GADD153-mediated pathway. Eur J Pharmacol. 2007 Feb 5;556(1-3):45-54. Epub 2006 Nov 3.

    Yeh TC, Chiang PC, Li TK, Hsu JL, Lin CJ, Wang SW, Peng CY, Guh JH. Genistein induces apoptosis in human hepatocellular carcinomas via interaction of endoplasmic reticulum stress and mitochondrial insult. Biochem Pharmacol. 2007 Mar 15;73(6):782-92. Epub 2006 Dec 3.

    Bani-Yaghoub M, Kubu CJ, Cowling R, Rochira J, Nikopoulos GN, Bellum S, Verdi JM. A switch in numb isoforms is a critical step in cortical development. Dev Dyn. 2007 Jan 25; [Epub ahead of print]

    Du L, Pollard JM, Gatti RA. Correction of prototypic ATM splicing mutations and aberrant ATM function with antisense morpholino oligonucleotides. Proc Natl Acad Sci U S A. 2007 Mar 26; [Epub ahead of print]

    Chow G, Knudson CB, Knudson W. Expression and cellular localization of human hyaluronidase-2 in articular chondrocytes and cultured cell lines. Osteoarthritis Cartilage. 2006 Sep;14(9):849-58.

    Sample R, Bryan L, Long S, Majji S, Hoskins G, Sinning A, Olivier J, Chinchar VG. Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II. Virology. 2007 Feb 20;358(2):311-20. Epub 2006 Oct 3.

    Isenberg JS, Romeo MJ, Abu-Asab M, Tsokos M, Oldenborg A, Pappan L, Wink DA, Frazier WA, Roberts DD. Increasing Survival of Ischemic Tissue by Targeting CD47. Circ Res. 2007 Feb 9; [Epub ahead of print]

    Mironov S.ADP regulates movements of mitochondria in neurons. Biophys J. 2007 Feb 2; [Epub ahead of print]

    Hao Q, Hong SH, Maret W.Lipid raft-dependent endocytosis of metallothionein in HepG2 cells. J Cell Physiol. 2007 Feb;210(2):428-35.

    Madhavan M, Haynes TL, Frisch NC, Call MK, Minich CM, Tsonis PA, Del Rio-Tsonis K. The role of Pax-6 in lens regeneration. Proc Natl Acad Sci U S A. 2006 Oct 3;103(40):14848-53. Epub 2006 Sep 26.

    Dunlap KA, Palmarini M, Varela M, Burghardt RC, Hayashi K, Farmer JL, Spencer TE. Endogenous retroviruses regulate periimplantation placental growth and differentiation. Proc Natl Acad Sci U S A. 2006 Sep 26;103(39):14390-5. Epub 2006 Sep 15.

    Sands WA, Woolson HD, Milne GR, Rutherford C, Palmer TM. Exchange Protein Activated by Cyclic AMP (Epac)-Mediated Induction of Suppressor of Cytokine Signaling 3 (SOCS-3) in Vascular Endothelial Cells. Mol Cell Biol. 2006 Sep;26(17):6333-46.

    Chih B, Gollan L, Scheiffele P. Alternative splicing controls selective trans-synaptic interactions of the neuroligin-neurexin complex. Neuron. 2006 Jul 20;51(2):171-8.

    Talbot L, Gonzalez P, Liton P, Epstein DL, Challa P. Optimizing Delivery of Novel Anti-Sense Oligonucleotides to Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci., May 2006; 47: 1874.

    Choudhury A, Marks DL, Proctor KM, Gould GW, Pagano RE. Regulation of caveolar endocytosis by syntaxin 6-dependent delivery of membrane components to the cell surface. Nat Cell Biol. 2006 Apr;8(4):317-28. Epub 2006 Mar 26.

    Boyden ED, Dietrich WF. Nalp1b controls mouse macrophage susceptibility to anthrax lethal toxin. Nat Genet. 2006 Feb;38(2):240-4. Epub 2006 Jan 22.

    Alison J. Tyson-Capper and G. Nicholas Europe-Finner. Novel Targeting of COX-2 pre-mRNA Using Antisense Morpholino Oligonucleotides Directed To The 3' Acceptor And 5' Donor Splice Sites Of Exon 4: Suppression Of COX-2 Activity In Human Amnion-Derived WISH And Myometrial Cells. Mol Pharmacol. 2006 Mar;69(3):796-804. Epub 2005 Dec 14.

    Masaki M et al. Smad1 protects cardiomyocytes from ischemia-reperfusion injury. Circulation. 2005 May 31;111(21):2752-9. Epub 2005 May 23.

    Dunlap K, Hayashi K, Adelson D, Palmarini M, Spencer T. Ovine Endogenous Betaretroviruses (enJSRVs) Regulate Conceptus Growth and Development. Platform presentation at the Society for the Study of Reproduction 2005 meeting, Quebec, Canada.

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