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Poly-Arginine-HA Tagged Reprogramming Factors Description Poly-arginine-HA tagged reprogramming factors allow the production of recombinant cell-penetrating reprogramming factors. They correspond to the four transcription factors, Oct4, Sox2, Klf4, and c-Myc (OSKM), fused at their C terminus to a poly-arginine (i.e. 11R) peptide in tandem with 3 motifs of the hemaglutinine (HA) tag. The poly-arginine peptide enables the recombinant proteins to readily enter the cells and have been shown to allow their translocation into the nucleus 1, 2 . The HA tag is useful for their detection by Western blot or their purification by affinity chromatography. Poly-arginine-HA tagged reprogramming factors are cloned in the pUNO1 plasmid within a mammalian expression cassette comprising the EF-1a/HTLV composite promoter and the SV40 poly adenylation sequence. pUNO1 plasmids are selectable in E. coli and mammalian cells with blasticidin. Application Poly-arginine-HA tagged reprogramming factors are designed for the generation of protein-induced pluripotent cells. Following their transfection into mammalian cell lines, such as HEK293, the cell extracts can be used crude to treat the target cells or processed to purifiy the poly-arginine-HA-tagged proteins on an anti-HA affinity column. Contents and Storage Each pUNO1 plasmid is provided as a lyophilized transformed E. coli strain on a paper disk. Transformed strains are shipped at room temperature and should be stored at -20°C. Each pUNO1 is provided with 4 pouches of E. coli Fast-Media ® Blas (2 TB and 2 Agar). For more information about Fast-Media ® , see pages 44-45. Reprogramming Enhancers 1. Zhou H. et al., 2009. Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell. 4(5):381-4. 2. Kim D. et al., 2009. Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. Cell Stem Cell. 4(6):472-6. PRODUCT CAT. CODE (HUMAN) CAT. CODE (MOUSE) pUNO1-OCT4-11RHA puno1rha-hoct4 puno1rha-moct4 pUNO1-SOX2-11RHA puno1rha-hsox2 puno1rha-msox2 pUNO1-KLF4-11RHA puno1rha-hklf4 puno1rha-mklf4 pUNO1-cMYC-11RHA puno1rha-hmycb puno1rha-mmycb InvivoGen offers a selection of small molecules that have recently been reported to either enhance reprogramming efficiencies or substitute for specific reprogramming factors. Among these molecules, some are known to affect chromatin modifications while others influence signal transduction pathways. PRODUCT DESCRIPTION WORKING CONCENTRATION QUANTITY CATALOG CODE REF. INFO 5-Aza-cytidine DNA methyltransferase inhibitor 2 μM 100 mg inh-aza 1, 2 p 157 Bix-01294 G9a histone methyltransferase inhibitor 1 μM 2 mg inh-bix 3, 4 p 157 PD0325901 MEK inhibitor 0.5 μM 2 mg inh-pd32 5, 6 p 156 SB431542 TGF-b receptor inhibitor 2 μM 5 mg inh-sb43 6, 7 p 155 Valproic Acid HDAC inhibitor 2 mM 5 g inh-vpa 2, 8 p 157 1. Mikkelsen TS. et al., 2008. Dissecting direct reprogramming through integrative genomic analysis. Nature. 454(7200):49-55. 2. Huangfu D. et al., 2008a. Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds. Nat Biotechnol. 26(7):795-7. 3. Epsztejn-Litman S. et al., 2008. De novo DNA methylation promoted by G9a prevents reprogramming of embryonically silenced genes. Nat Struct Mol Biol. 15(11):1176-83. 4. Shi Y et al., 2008. A combined chemical and genetic approach for the generation of induced pluripotent stem cells. Cell Stem Cell. 2(6):525-8. 5. Ying QL. et al., 2008. The ground state of embryonic stem cell self-renewal. Nature. 453(7194):519-23. 6. Lin T. et al., 2009. A chemical platform for improved induction of human iPSCs. Nat Methods. 6(11):805-8. 7. IInman GJ. et al., 2002. SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol. 2002 Jul;62(1):65-74. 8 . Huangfu D. et al., 2008b. Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol. 26(11):1269-75. Contents and Storage www.invivogen.com/ipsc 3xHA 11R Each product is provided as a solid and shipped at room temperature. Store at room temperature, 4°C or -20°C according to the product label. 21 CELL CULTURE & TRANSFECTION 1
2 MAMMALIAN EXPRESSION VECTORS Lentiviral Vectors 23 Cloning Vectors 26 Genes - Open Reading Frames 36 Cellular Promoters 38 E. coli Growth & Selection 42
Page 2 and 3: INNOVATION WITHIN REACH InvivoGen
Page 4 and 5: CONTENTS AT A GLANCE 1 CELL CULTURE
Page 6 and 7: 4 CYTOKINE SIGNALING TABLE of CONTE
Page 8 and 9: Mycoplasma Detection & Elimination
Page 10 and 11: PlasmoTest Contents - HEK-Blue -2
Page 12 and 13: Primocin - The Antimicrobial Shiel
Page 14 and 15: SEAP Reporter Gene System Secreted
Page 16 and 17: HEK-Blue Detection - Real-Time Det
Page 18 and 19: Blasticidin S Description Blasticid
Page 20 and 21: LyoVec New formulation LyoVec is
Page 24 and 25: LENTI-Smart - Lentiviral Vector Pr
Page 26 and 27: Lentiviral Vector Production and Ce
Page 28 and 29: Antibody generation using pFUSE-CHI
Page 30 and 31: Engineering Fc regions for altered
Page 32 and 33: pBROAD & pWHERE - Optimized Vector
Page 34 and 35: pSELECT-Tag - Expression of a GFP-,
Page 36 and 37: pVIVO - Expression of Two Genes of
Page 38 and 39: pORF Description pORF plasmids feat
Page 40 and 41: Prom A-List - Human and Rodent Pro
Page 42 and 43: PROMOTER GENE TISSUE DISTRIBUTION S
Page 44 and 45: ChemiComp E. coli ChemiComp E. coli
Page 46 and 47: Fast-Media ® Preparation 1. Empty
Page 48 and 49: RIG-I-like Receptors 54 RLR & RLR-R
Page 50 and 51: Toll-Like Receptors Toll-Like Recep
Page 52 and 53: www.invivogen.com/innate-immunity 5
Page 54 and 55: IPAF and NAIP5 IPAF (NLRC4, CLAN/CA
Page 56 and 57: C-Type Lectin Receptors C-type lect
Page 58 and 59: pUNO pUNO-HA Toll-Like Receptors (T
Page 60 and 61: GENE NAME/ALIASES DESCRIPTION CAT.
Page 62 and 63: pDUO - Gene Associations Features a
Page 64 and 65: pZERO-TLR & pDeNy - Dominant Negati
Page 66 and 67: 293/TLR & 293/NOD Clones 293/TLR Cl
Page 68 and 69: Response of HEK-Blue hTLR2 cells t
Page 70 and 71: Ramos-Blue Cells - NF-kB/AP-1 Repo
Page 72 and 73:
MEF Cells C3H/TLR4mut & C3H/WT MEFs
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Antibodies for Detection ANTIBODY D
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- NF-kB binding site: Nuclear facto
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PRODUCT ORIGIN/DESCRIPTION TLR2 Ago
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PRODUCT ORIGIN/DESCRIPTION TLR9 Ago
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PRODUCT ORIGIN/DESCRIPTION Dectin-1
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LPS-PG (Porphyromonas gingivalis) R
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• FLA-ST ultrapure was purified b
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1. Zhao H. et al., 2004. Contributi
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1. Hornung V. et al., 2006. 5'-Trip
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TLR/NOD Ligand Screening InvivoGen
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Signal Transduction Inhibitors Invi
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Pepinh-MYD - MyD88 inhibitory pepti
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1. Martinon F,. et al., 2002. The i
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Nigericin - NLRP3 Inflammasome Indu
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Autophagy is a pathway by which cyt
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4 CYTOKINE SIGNALING Cytokine Genes
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Blue Cytokine Reporter Cells Blue
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HEK-Blue IFN-a/b Cells - Human Typ
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HEK-Blue IL-6 Cells - IL6 Reporter
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HEK-Blue IL-1b Cells - IL-1b Repor
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HEK-Blue IL-33/IL-1b Cells - IL-33
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HEK-Blue CD40L Cells - CD40L Repor
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5 ANTIBODIES & VACCINATION Antibodi
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Antibody generation using pFUSE-CHI
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Immunoglobulin A The mucosal surfac
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IgA Detection and Quantification In
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IgG Antibodies InvivoGen offers a s
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Mediators? 1. Leroux-Roels G., 2010
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Poly(I:C) Synthetic double-stranded
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pVAC - Induction of Neutralizing An
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CpG-Free DNA CpGs and the Immune Re
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pCpGfree-basic & pCpGfree-promoter
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pCpGfree-siRNA & pCpGfree-siRNADUO
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Custom-Made CpG-Free Genes InvivoGe
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RNA INTERFERENCE 7 psiRNA System I
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RNA INTERFERENCE 7 psiRNA-h7SK - 7S
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RNA INTERFERENCE 7 psiTEST System
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RNA INTERFERENCE 7 Ready-Made psiRN
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RNA INTERFERENCE 7 Custom-Made psiR
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INHIBITORS 8 Inhibitors PRODUCT DES
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INHIBITORS 8 Pattern Recognition Re
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INHIBITORS 8 Triptolide - NF-kB Inh
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INHIBITORS 8 Heat Shock Protein Inh
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9 CUSTOM SERVICES TLR/NOD Ligand Sc
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CUSTOM SERVICES 9 Custom Cloning :
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CUSTOM SERVICES 9 Prices 164 TERMS
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166 PRODUCT (QUANTITY) CAT. CODE PA
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A 17-AAG 17-AAGH2 17-AEP-GA 2-Amino
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- ODN 4084-F - ODN INH-1 - ODN INH-
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ARGENTINA Genbiotech S.R.L. Tel: +5
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