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plasmids pep4 e02s lm2l  (Addgene inc)


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    Addgene inc plasmids pep4 e02s lm2l
    Plasmids Pep4 E02s Lm2l, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcep4+m2l/pCEP4-M2L+(Plasmid+%2320926)/bio_rxiv__64898__2026__03__17__712482-84-30-34
    Average 93 stars, based on 22 article reviews
    plasmids pep4 e02s lm2l - by Bioz Stars, 2026-10
    93/100 stars

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    Transfection:

    Article Title: Generation of two induced pluripotent stem cell (iPSC) lines (HZSMHCI001-A and HZSMHCI004-A) from a mother-child dyad with major depressive disorder and bipolar disorder.
    Article Snippet: The resulting CD34+enriched cell population was subsequently reprogrammed by electroporation using a Lonza Nucleofector with the Nucleofector kit (Lonza, VPA-1003). .. Cells were transfected with three non-integrating episomal plasmids: pEP4EO2SEN2K, pEP4EO2SET2K, and pCEP4-M2L (Addgene, 20925, 20927, 20926), which together express OCT4, SOX2, NANOG, LIN28, c-MYC, KLF4, and SV40LT. .. Transfected cells were plated on Matrigel-coated plates (Corning, 354277) in Essential 8 medium (Thermo Fisher Scientific, A1517001).

    Article Title: Modeling high-risk Wilms tumors enables the discovery of therapeutic vulnerability.
    Article Snippet: .. Cellular reprogramming via episomal vector transfection Episomal vectors, including pEP4 E02S EN2K (plasmid #20925), pCEP4-M2L (#20926), and pEP4 E02S ET2K (#20927) supplied by Addgene (Cambridge, MA, USA) and gifted by Dr. James Thomson, were utilized as detailed in a prior publication.55 To createWiT49PRCs, WiT49 cells underwent transfection with a mix of these episomal plasmids carrying OCT4, SOX2, KLF4, c-MYC, NANOG, and LIN28A (3 mg of pEP4 E02S ET2K, 3 mg of pEP4 E02S EN2K, and 2 mg of pCEP4-M2L), following our established protocol.28 This process involved using the Nucleofector II device with the U-20 program (Amaxa, Walkersville, MD, USA) and the Nucleofector kit R (Lonza, Hayward, CA, USA). .. Post-transfection, cells were seeded in a Matrigel-coated 6-well plate with Essential 8 (E8) Medium (A1517001) (Invitrogen, Carlsbad, CA, USA), with daily medium changes to E7 (E8minus TGFB1) upon reaching 20–30% confluence.

    Plasmid Preparation:

    Article Title: Modeling high-risk Wilms tumors enables the discovery of therapeutic vulnerability
    Article Snippet: .. Episomal vectors, including pEP4 E02S EN2K (plasmid #20925), pCEP4-M2L (#20926), and pEP4 E02S ET2K (#20927) supplied by Addgene (Cambridge, MA, USA) and gifted by Dr. James Thomson, were utilized as detailed in a prior publication. ..

    Article Title: Modeling high-risk Wilms tumors enables the discovery of therapeutic vulnerability.
    Article Snippet: .. Cellular reprogramming via episomal vector transfection Episomal vectors, including pEP4 E02S EN2K (plasmid #20925), pCEP4-M2L (#20926), and pEP4 E02S ET2K (#20927) supplied by Addgene (Cambridge, MA, USA) and gifted by Dr. James Thomson, were utilized as detailed in a prior publication.55 To createWiT49PRCs, WiT49 cells underwent transfection with a mix of these episomal plasmids carrying OCT4, SOX2, KLF4, c-MYC, NANOG, and LIN28A (3 mg of pEP4 E02S ET2K, 3 mg of pEP4 E02S EN2K, and 2 mg of pCEP4-M2L), following our established protocol.28 This process involved using the Nucleofector II device with the U-20 program (Amaxa, Walkersville, MD, USA) and the Nucleofector kit R (Lonza, Hayward, CA, USA). .. Post-transfection, cells were seeded in a Matrigel-coated 6-well plate with Essential 8 (E8) Medium (A1517001) (Invitrogen, Carlsbad, CA, USA), with daily medium changes to E7 (E8minus TGFB1) upon reaching 20–30% confluence.

    Article Title: Human blood brain barrier model
    Article Snippet: .. Electroporation: 5.0×105 amniotic fluid (AF) cells were electroporated (nucleofected) with 1.3 pg of each plasmid (pEP4 E02S EN2K (Addgene—20925), pCEP4 M2L (Addgene—20926) and pEP4 E02S ET2K (Addgene—20927)), using the NucleofectorTM II Device Amaxa and the Basic Epithelial cell NucleofectorTM Kit (Lonza Cat #VPI 1005), using programs O-03 and 5.0×105 cells electroporated with program 1-03. .. Electroporated cells were plated on a Matrigel-coated 6 well plate in DME+20% FBS and 10 μM Y27632 Stemolecule (Stemgent).

    Article Title: Efficient derivation of transgene-free porcine induced pluripotent stem cells enables in vitro modeling of species-specific developmental timing
    Article Snippet: To 155 initiate a reprogramming experiment, primary fibroblasts were lifted by TrypLE Express Enzyme 156 (TrypLE, ThermoFisher Scientific Cat. No. 12605028), counted, and electroporation carried out 157 using the Neon Transfection System (ThermoFisher Scientific Cat. No. MPK1096) with program 158 20 (pulse voltage: 1150; pulse width: 30; pulse number: 2). .. In each round of the electroporation, 159 all 1.0 × 105 cells were electroporated with a total of 2.0μg plasmid DNA, consisting of pEP4-160 E02SEN2K (0.5μg), pCEP4-M2L (0.5μg), pEP4-E02SET2K (0.5μg) and pSimple-miRNA302/367 161 (0.5μg, Addgene plasmid # 98748), combined with 0.5ug of EBNA1 mRNA in vitro transcribed 162 from the template Sp6-EBNA1 (Addgene plasmid # 98749). .. These plasmids were kind gifts from 163 Drs. James Thomson, Sara Howden, and Jim Vadolas (Addgene plasmid # 20925, 20926, 20927, 164 98748 and 98749)(Howden et al., 2015; Howden et al., 2006; Yu et al., 2009).

    Article Title: Establishment of Transgene-Free Porcine Induced Pluripotent Stem Cells.
    Article Snippet: .. REAGENTS AND SOLUTIONS Blocking buffer 500 ml PBS (Thermo Fisher Scientific, cat. no. 14190250) [97% (v/v) final] 10 ml bovine serum albumin (BSA; Sigma, cat. no. A7030-100G) [2% (v/v) final] 5 ml fetal bovine serum (FBS), heat inactivated (Thermo Fisher Scientific, cat. no. SH30396.03) [1% (v/v) final] Store ≤6 months at 4°C Episomal plasmid mix pEP4-E02SEN2K (Addgene plasmid # 20925) pCEP4-M2L (Addgene plasmid # 20926) pEP4-E02SET2K (Addgene plasmid # 20927) Precipitate and concentrate at 1:1:1 ratio, aiming for final concentration of 1.0 μg/μl Store at 4°C if in use or freeze at −20°C for long-term storage Fibroblast freezing medium 5 ml dimethyl sulfoxide (DMSO; Sigma, cat. no. D2650-100ML) [10% (v/v) final] 20 ml fibroblast medium (see recipe) [40% (v/v) final] Conrad et al. 15 of 22 Current Protocols 26911299, 2024, 5, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpz1.1012 by N at Prov Indonesia, W iley O nline L ibrary on [23/01/2026]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense PBS/FBS 500 ml PBS (Thermo Fisher Scientific, cat. no. 14190250) [99% (v/v) final] 5 ml FBS, stem cell qualified (Sigma, cat. no. ES-009-B) [1% (v/v) final] Prepare in biosafety cabinet and filter (0.22 μm; 250- and 500-ml 0.22-μm filters, Sigma, cat. no. S2GPU02RE and S2GPU05RE) before use Store ≤2 months at 4°C COMMENTARY Background Information The genesis of porcine reprogramming can be traced back to 2009, when multiple groups reported the successful reprogramming of porcine somatic cells by integrating exogenous reprogramming factors into the host cell genome using viral vectors (Esteban et al., 2009; Ezashi et al., 2009; Wu et al., 2009).

    Electroporation:

    Article Title: Human blood brain barrier model
    Article Snippet: .. Electroporation: 5.0×105 amniotic fluid (AF) cells were electroporated (nucleofected) with 1.3 pg of each plasmid (pEP4 E02S EN2K (Addgene—20925), pCEP4 M2L (Addgene—20926) and pEP4 E02S ET2K (Addgene—20927)), using the NucleofectorTM II Device Amaxa and the Basic Epithelial cell NucleofectorTM Kit (Lonza Cat #VPI 1005), using programs O-03 and 5.0×105 cells electroporated with program 1-03. .. Electroporated cells were plated on a Matrigel-coated 6 well plate in DME+20% FBS and 10 μM Y27632 Stemolecule (Stemgent).

    Article Title: Efficient derivation of transgene-free porcine induced pluripotent stem cells enables in vitro modeling of species-specific developmental timing
    Article Snippet: To 155 initiate a reprogramming experiment, primary fibroblasts were lifted by TrypLE Express Enzyme 156 (TrypLE, ThermoFisher Scientific Cat. No. 12605028), counted, and electroporation carried out 157 using the Neon Transfection System (ThermoFisher Scientific Cat. No. MPK1096) with program 158 20 (pulse voltage: 1150; pulse width: 30; pulse number: 2). .. In each round of the electroporation, 159 all 1.0 × 105 cells were electroporated with a total of 2.0μg plasmid DNA, consisting of pEP4-160 E02SEN2K (0.5μg), pCEP4-M2L (0.5μg), pEP4-E02SET2K (0.5μg) and pSimple-miRNA302/367 161 (0.5μg, Addgene plasmid # 98748), combined with 0.5ug of EBNA1 mRNA in vitro transcribed 162 from the template Sp6-EBNA1 (Addgene plasmid # 98749). .. These plasmids were kind gifts from 163 Drs. James Thomson, Sara Howden, and Jim Vadolas (Addgene plasmid # 20925, 20926, 20927, 164 98748 and 98749)(Howden et al., 2015; Howden et al., 2006; Yu et al., 2009).

    In Vitro:

    Article Title: Efficient derivation of transgene-free porcine induced pluripotent stem cells enables in vitro modeling of species-specific developmental timing
    Article Snippet: To 155 initiate a reprogramming experiment, primary fibroblasts were lifted by TrypLE Express Enzyme 156 (TrypLE, ThermoFisher Scientific Cat. No. 12605028), counted, and electroporation carried out 157 using the Neon Transfection System (ThermoFisher Scientific Cat. No. MPK1096) with program 158 20 (pulse voltage: 1150; pulse width: 30; pulse number: 2). .. In each round of the electroporation, 159 all 1.0 × 105 cells were electroporated with a total of 2.0μg plasmid DNA, consisting of pEP4-160 E02SEN2K (0.5μg), pCEP4-M2L (0.5μg), pEP4-E02SET2K (0.5μg) and pSimple-miRNA302/367 161 (0.5μg, Addgene plasmid # 98748), combined with 0.5ug of EBNA1 mRNA in vitro transcribed 162 from the template Sp6-EBNA1 (Addgene plasmid # 98749). .. These plasmids were kind gifts from 163 Drs. James Thomson, Sara Howden, and Jim Vadolas (Addgene plasmid # 20925, 20926, 20927, 164 98748 and 98749)(Howden et al., 2015; Howden et al., 2006; Yu et al., 2009).

    Clone Assay:

    Article Title: A familiar cerebral cavernous malformations (FCCMs) patient with CCM1 deletion-frameshift mutation generates an induced pluripotent stem cell line (NFHHMUi001-A).
    Article Snippet: As instructed, Nucleofector-2b-Device, CD34+ Nucleofector Kit (Lonza) and program T-16 were applied to transfect 1 × 106 PBMCs with episomal iPSC reprogramming plasmids. .. The human transgenes OCT4, SOX2, NANOG, LIN28, c-MYC, KLF4 and SV40LT were cloned into the following plasmids: pEP4EO2SEN2K (addgene#20925), pCEP4-M2L (addgene#20926), pEP4EO2SET2K (addgene#20927). ..

    Blocking Assay:

    Article Title: Establishment of Transgene-Free Porcine Induced Pluripotent Stem Cells.
    Article Snippet: .. REAGENTS AND SOLUTIONS Blocking buffer 500 ml PBS (Thermo Fisher Scientific, cat. no. 14190250) [97% (v/v) final] 10 ml bovine serum albumin (BSA; Sigma, cat. no. A7030-100G) [2% (v/v) final] 5 ml fetal bovine serum (FBS), heat inactivated (Thermo Fisher Scientific, cat. no. SH30396.03) [1% (v/v) final] Store ≤6 months at 4°C Episomal plasmid mix pEP4-E02SEN2K (Addgene plasmid # 20925) pCEP4-M2L (Addgene plasmid # 20926) pEP4-E02SET2K (Addgene plasmid # 20927) Precipitate and concentrate at 1:1:1 ratio, aiming for final concentration of 1.0 μg/μl Store at 4°C if in use or freeze at −20°C for long-term storage Fibroblast freezing medium 5 ml dimethyl sulfoxide (DMSO; Sigma, cat. no. D2650-100ML) [10% (v/v) final] 20 ml fibroblast medium (see recipe) [40% (v/v) final] Conrad et al. 15 of 22 Current Protocols 26911299, 2024, 5, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpz1.1012 by N at Prov Indonesia, W iley O nline L ibrary on [23/01/2026]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense PBS/FBS 500 ml PBS (Thermo Fisher Scientific, cat. no. 14190250) [99% (v/v) final] 5 ml FBS, stem cell qualified (Sigma, cat. no. ES-009-B) [1% (v/v) final] Prepare in biosafety cabinet and filter (0.22 μm; 250- and 500-ml 0.22-μm filters, Sigma, cat. no. S2GPU02RE and S2GPU05RE) before use Store ≤2 months at 4°C COMMENTARY Background Information The genesis of porcine reprogramming can be traced back to 2009, when multiple groups reported the successful reprogramming of porcine somatic cells by integrating exogenous reprogramming factors into the host cell genome using viral vectors (Esteban et al., 2009; Ezashi et al., 2009; Wu et al., 2009).

    Concentration Assay:

    Article Title: Establishment of Transgene-Free Porcine Induced Pluripotent Stem Cells.
    Article Snippet: .. REAGENTS AND SOLUTIONS Blocking buffer 500 ml PBS (Thermo Fisher Scientific, cat. no. 14190250) [97% (v/v) final] 10 ml bovine serum albumin (BSA; Sigma, cat. no. A7030-100G) [2% (v/v) final] 5 ml fetal bovine serum (FBS), heat inactivated (Thermo Fisher Scientific, cat. no. SH30396.03) [1% (v/v) final] Store ≤6 months at 4°C Episomal plasmid mix pEP4-E02SEN2K (Addgene plasmid # 20925) pCEP4-M2L (Addgene plasmid # 20926) pEP4-E02SET2K (Addgene plasmid # 20927) Precipitate and concentrate at 1:1:1 ratio, aiming for final concentration of 1.0 μg/μl Store at 4°C if in use or freeze at −20°C for long-term storage Fibroblast freezing medium 5 ml dimethyl sulfoxide (DMSO; Sigma, cat. no. D2650-100ML) [10% (v/v) final] 20 ml fibroblast medium (see recipe) [40% (v/v) final] Conrad et al. 15 of 22 Current Protocols 26911299, 2024, 5, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpz1.1012 by N at Prov Indonesia, W iley O nline L ibrary on [23/01/2026]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense PBS/FBS 500 ml PBS (Thermo Fisher Scientific, cat. no. 14190250) [99% (v/v) final] 5 ml FBS, stem cell qualified (Sigma, cat. no. ES-009-B) [1% (v/v) final] Prepare in biosafety cabinet and filter (0.22 μm; 250- and 500-ml 0.22-μm filters, Sigma, cat. no. S2GPU02RE and S2GPU05RE) before use Store ≤2 months at 4°C COMMENTARY Background Information The genesis of porcine reprogramming can be traced back to 2009, when multiple groups reported the successful reprogramming of porcine somatic cells by integrating exogenous reprogramming factors into the host cell genome using viral vectors (Esteban et al., 2009; Ezashi et al., 2009; Wu et al., 2009).



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