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Endo-PorterEndo-Porter simple delivery reagent
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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) 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.
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 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] 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. 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. 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 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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