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pcdna3 apex2 nes  (Addgene inc)


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    Structured Review

    Addgene inc pcdna3 apex2 nes
    Pcdna3 Apex2 Nes, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna3+apex2+nes/pcDNA3+APEX2-NES+(Plasmid+%2349386)/pm41807954-183-1-3
    Average 95 stars, based on 114 article reviews
    pcdna3 apex2 nes - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    other:

    Article Title: The Kinase CK1α coordinates Initiation and Termination of the cGAS-STING Pathway
    Article Snippet: nLC-MS/MS analyses were performed on a nanoElute 2 nanoflow system coupled to a TIMS-TOF Pro 2 mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany).

    Plasmid Preparation:

    Article Title: Kremen1 dependence receptor induces SEC24C and ATG9A-dependent cell death.
    Article Snippet: We cloned the Kremen1 sequence into the inducible Sleeping Beauty transposon system into the Psbtet backone (Addgene #60495 by infusion cloning (In-Fusion HD Cloning Kit, Takara bio 639649). .. The pcDNA3 APEX2-NES (addgene #49386) was used to fuse the APEX2 enzyme at the C-terminus of Kremen1 in the Psbtet plasmid using infusion cloning. .. The human breast cancer cell line MDA-MB231, human pancreatic cancer cell line BxPC3 and human embryonic kidney HEK293T cells were obtained from ATCC (UK) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin.

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells
    Article Snippet: .. As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene. .. Human Umbilical Vein Endothelial Cells (Lonza, pooled, Cat#C2519A) were cultured in Endothelial Cell Basal Media 2 (Promocell) supplemented with Growth Medium 2 supplement mix (composition in Supp.

    Article Title: Ribosome-binding protein 1 maintains peroxisome biogenesis.
    Article Snippet: .. A FLAG–APEX2 fragment was amplified from pcDNA3 APEX2-NES (Addgene plasmid 49386; Lam et al., 2015), ePTS1 was generated using corresponding primers. .. The resulting fused construct was inserted into pLL3.7 vector (Addgene plasmid 11795; Rubinson et al., 2003) using EcoRI and NheI restriction sites.

    Article Title: A proximity labeling approach to identify proteins that associate with synaptonemal complex components in Drosophila melanogaster females
    Article Snippet: .. APEX2 was PCR amplified from pcDNA3 APEX2-NES (Addgene Plasmid #49386) without both the stop codon and nuclear export signal with NotI and NdeI restriction sites. ..

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells
    Article Snippet: To target HUVEC, a lentiviral variant without mKO2, due to size restrictions, was made by PCRing JAG1-APEX2 with the primers FW:TATTATCTAGATTATTTGTCATCATCGTCCTTATAATCGGCG and RV:CGAATGGAGTACATCGTAGGGGATCC, and cloned with XbaI and BamhI into pENTR1a (addgene #19364), and subsequently transferred to pLenti-DEST (Addgene #19068) using gateway recombination (LR-clonase 2). .. As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal (NES) for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene. ..

    Cloning:

    Article Title: Kremen1 dependence receptor induces SEC24C and ATG9A-dependent cell death.
    Article Snippet: We cloned the Kremen1 sequence into the inducible Sleeping Beauty transposon system into the Psbtet backone (Addgene #60495 by infusion cloning (In-Fusion HD Cloning Kit, Takara bio 639649). .. The pcDNA3 APEX2-NES (addgene #49386) was used to fuse the APEX2 enzyme at the C-terminus of Kremen1 in the Psbtet plasmid using infusion cloning. .. The human breast cancer cell line MDA-MB231, human pancreatic cancer cell line BxPC3 and human embryonic kidney HEK293T cells were obtained from ATCC (UK) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin.

    Control:

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells
    Article Snippet: .. As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene. .. Human Umbilical Vein Endothelial Cells (Lonza, pooled, Cat#C2519A) were cultured in Endothelial Cell Basal Media 2 (Promocell) supplemented with Growth Medium 2 supplement mix (composition in Supp.

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells
    Article Snippet: To target HUVEC, a lentiviral variant without mKO2, due to size restrictions, was made by PCRing JAG1-APEX2 with the primers FW:TATTATCTAGATTATTTGTCATCATCGTCCTTATAATCGGCG and RV:CGAATGGAGTACATCGTAGGGGATCC, and cloned with XbaI and BamhI into pENTR1a (addgene #19364), and subsequently transferred to pLenti-DEST (Addgene #19068) using gateway recombination (LR-clonase 2). .. As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal (NES) for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene. ..

    Labeling:

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells
    Article Snippet: .. As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene. .. Human Umbilical Vein Endothelial Cells (Lonza, pooled, Cat#C2519A) were cultured in Endothelial Cell Basal Media 2 (Promocell) supplemented with Growth Medium 2 supplement mix (composition in Supp.

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells
    Article Snippet: To target HUVEC, a lentiviral variant without mKO2, due to size restrictions, was made by PCRing JAG1-APEX2 with the primers FW:TATTATCTAGATTATTTGTCATCATCGTCCTTATAATCGGCG and RV:CGAATGGAGTACATCGTAGGGGATCC, and cloned with XbaI and BamhI into pENTR1a (addgene #19364), and subsequently transferred to pLenti-DEST (Addgene #19068) using gateway recombination (LR-clonase 2). .. As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal (NES) for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene. ..

    Construct:

    Article Title: Concerted remodelling of the postsynaptic spine and RNA granule by cLTP
    Article Snippet: Animal experimental procedures were approved by the ethics committee of the National Research Council of Spain (Approval number: B9900083). .. Lentiviral constructs pLKO-RFP-shCtrl (Addgene, 69040) and pLKO-RFP-shDbn1 containing the shRNA sequence GCAGTCTATCTTTGGTGACCA (TRCN0000090077) against the Dbn1 gene were used in knockdown experiments. pFUW-FLAG-APEX2, pFUW-FLAG-APEX2-DBN1 and pFUW-FLAG-APEX2-IGF2BP1 were used to perform accessibility and proximity assays, and derived from pcDNA3 APEX2-NES (Addgene, 49386) and pFUW (Addgene, 14882). .. Dbn1, Dbn1 ΔIDR and Igf2bp1 were amplified using Phusion High-Fidelity DNA Polymerase (Thermo Fisher, 10024537) from mouse cDNAs. mEGFP-C3 (Clontech) and mScarlet-C1 (Addgene, 85044) were used as host vectors.

    shRNA:

    Article Title: Concerted remodelling of the postsynaptic spine and RNA granule by cLTP
    Article Snippet: Animal experimental procedures were approved by the ethics committee of the National Research Council of Spain (Approval number: B9900083). .. Lentiviral constructs pLKO-RFP-shCtrl (Addgene, 69040) and pLKO-RFP-shDbn1 containing the shRNA sequence GCAGTCTATCTTTGGTGACCA (TRCN0000090077) against the Dbn1 gene were used in knockdown experiments. pFUW-FLAG-APEX2, pFUW-FLAG-APEX2-DBN1 and pFUW-FLAG-APEX2-IGF2BP1 were used to perform accessibility and proximity assays, and derived from pcDNA3 APEX2-NES (Addgene, 49386) and pFUW (Addgene, 14882). .. Dbn1, Dbn1 ΔIDR and Igf2bp1 were amplified using Phusion High-Fidelity DNA Polymerase (Thermo Fisher, 10024537) from mouse cDNAs. mEGFP-C3 (Clontech) and mScarlet-C1 (Addgene, 85044) were used as host vectors.

    Sequencing:

    Article Title: Concerted remodelling of the postsynaptic spine and RNA granule by cLTP
    Article Snippet: Animal experimental procedures were approved by the ethics committee of the National Research Council of Spain (Approval number: B9900083). .. Lentiviral constructs pLKO-RFP-shCtrl (Addgene, 69040) and pLKO-RFP-shDbn1 containing the shRNA sequence GCAGTCTATCTTTGGTGACCA (TRCN0000090077) against the Dbn1 gene were used in knockdown experiments. pFUW-FLAG-APEX2, pFUW-FLAG-APEX2-DBN1 and pFUW-FLAG-APEX2-IGF2BP1 were used to perform accessibility and proximity assays, and derived from pcDNA3 APEX2-NES (Addgene, 49386) and pFUW (Addgene, 14882). .. Dbn1, Dbn1 ΔIDR and Igf2bp1 were amplified using Phusion High-Fidelity DNA Polymerase (Thermo Fisher, 10024537) from mouse cDNAs. mEGFP-C3 (Clontech) and mScarlet-C1 (Addgene, 85044) were used as host vectors.

    Knockdown:

    Article Title: Concerted remodelling of the postsynaptic spine and RNA granule by cLTP
    Article Snippet: Animal experimental procedures were approved by the ethics committee of the National Research Council of Spain (Approval number: B9900083). .. Lentiviral constructs pLKO-RFP-shCtrl (Addgene, 69040) and pLKO-RFP-shDbn1 containing the shRNA sequence GCAGTCTATCTTTGGTGACCA (TRCN0000090077) against the Dbn1 gene were used in knockdown experiments. pFUW-FLAG-APEX2, pFUW-FLAG-APEX2-DBN1 and pFUW-FLAG-APEX2-IGF2BP1 were used to perform accessibility and proximity assays, and derived from pcDNA3 APEX2-NES (Addgene, 49386) and pFUW (Addgene, 14882). .. Dbn1, Dbn1 ΔIDR and Igf2bp1 were amplified using Phusion High-Fidelity DNA Polymerase (Thermo Fisher, 10024537) from mouse cDNAs. mEGFP-C3 (Clontech) and mScarlet-C1 (Addgene, 85044) were used as host vectors.

    Derivative Assay:

    Article Title: Concerted remodelling of the postsynaptic spine and RNA granule by cLTP
    Article Snippet: Animal experimental procedures were approved by the ethics committee of the National Research Council of Spain (Approval number: B9900083). .. Lentiviral constructs pLKO-RFP-shCtrl (Addgene, 69040) and pLKO-RFP-shDbn1 containing the shRNA sequence GCAGTCTATCTTTGGTGACCA (TRCN0000090077) against the Dbn1 gene were used in knockdown experiments. pFUW-FLAG-APEX2, pFUW-FLAG-APEX2-DBN1 and pFUW-FLAG-APEX2-IGF2BP1 were used to perform accessibility and proximity assays, and derived from pcDNA3 APEX2-NES (Addgene, 49386) and pFUW (Addgene, 14882). .. Dbn1, Dbn1 ΔIDR and Igf2bp1 were amplified using Phusion High-Fidelity DNA Polymerase (Thermo Fisher, 10024537) from mouse cDNAs. mEGFP-C3 (Clontech) and mScarlet-C1 (Addgene, 85044) were used as host vectors.

    Amplification:

    Article Title: Ribosome-binding protein 1 maintains peroxisome biogenesis.
    Article Snippet: .. A FLAG–APEX2 fragment was amplified from pcDNA3 APEX2-NES (Addgene plasmid 49386; Lam et al., 2015), ePTS1 was generated using corresponding primers. .. The resulting fused construct was inserted into pLL3.7 vector (Addgene plasmid 11795; Rubinson et al., 2003) using EcoRI and NheI restriction sites.

    Article Title: A proximity labeling approach to identify proteins that associate with synaptonemal complex components in Drosophila melanogaster females
    Article Snippet: .. APEX2 was PCR amplified from pcDNA3 APEX2-NES (Addgene Plasmid #49386) without both the stop codon and nuclear export signal with NotI and NdeI restriction sites. ..

    Generated:

    Article Title: Ribosome-binding protein 1 maintains peroxisome biogenesis.
    Article Snippet: .. A FLAG–APEX2 fragment was amplified from pcDNA3 APEX2-NES (Addgene plasmid 49386; Lam et al., 2015), ePTS1 was generated using corresponding primers. .. The resulting fused construct was inserted into pLL3.7 vector (Addgene plasmid 11795; Rubinson et al., 2003) using EcoRI and NheI restriction sites.

    Polymerase Chain Reaction:

    Article Title: A proximity labeling approach to identify proteins that associate with synaptonemal complex components in Drosophila melanogaster females
    Article Snippet: .. APEX2 was PCR amplified from pcDNA3 APEX2-NES (Addgene Plasmid #49386) without both the stop codon and nuclear export signal with NotI and NdeI restriction sites. ..



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    (A) Shown is the current model of the D. melanogaster SC. (B) Top view model of the D. melangaster SC with a schematic of the proximity ligation by <t>APEX2::C(2)M</t> and Cona::APEX2. APEX2::C(2)M and Cona::APEX2 incorporate into the LE and central region of the SC. In the presence of biotin-phenol and H2O2 APEX2 transfers biotin to electron-rich amino acids on nearby proteins. Nuclei are isolated from biotin-phenol treated ovaries and the biotinylated proteins were isolated on streptavidin coated beads for mass spectrometry analysis. Adapted from ( Cahoon et al . 2017 ).
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    Image Search Results


    Proximity biotin labeling, mass spectroscopy, and shear platform to identify shear stress-sensitive Jag1-interacting proteins (A) The engineered ascorbate peroxidase (APEX2) converts biotin tyramide to a free radical-carrying species in the presence H 2 O 2 , allowing labeling of nearby peptides. Coupling APEX2 to Jag1 and performing labeling interaction under different mechanical loading conditions allows the identification of mechanoresponsive Jagged-interacting factors (mJIFy and mJIFz). (B) Jag1 engineered with APEX2 on the C-terminal (intracellular) domain enables labeling of cytoplasmic proximal proteins in the presence or absence of flow-induced shear stress. (C) Immunoblot of Jag1 and Lenti Jag1-APEX2 expressions. HUVECs were transduced with Lenti-Jag1-APEX2, exposed to static or shear conditions, and blotted for Jag1 and Actin. (D and E) Immunofluorescence microscopy of cells labeled with streptavidin 488 (SA488, green) and 4',6-diamidino-2-phylindole (DAPI; blue). MCF7 cells were transduced with (D) Lenti-APEX2-NES or (E) Lenti-Jag1-APEX2, biotinylated, and stained with streptavidin 488 and DAPI; scale bars, 25 μm.

    Journal: iScience

    Article Title: Mechanosensitive interactions between Jag1 and Myo1c control Jag1 trafficking in endothelial cells

    doi: 10.1016/j.isci.2025.113879

    Figure Lengend Snippet: Proximity biotin labeling, mass spectroscopy, and shear platform to identify shear stress-sensitive Jag1-interacting proteins (A) The engineered ascorbate peroxidase (APEX2) converts biotin tyramide to a free radical-carrying species in the presence H 2 O 2 , allowing labeling of nearby peptides. Coupling APEX2 to Jag1 and performing labeling interaction under different mechanical loading conditions allows the identification of mechanoresponsive Jagged-interacting factors (mJIFy and mJIFz). (B) Jag1 engineered with APEX2 on the C-terminal (intracellular) domain enables labeling of cytoplasmic proximal proteins in the presence or absence of flow-induced shear stress. (C) Immunoblot of Jag1 and Lenti Jag1-APEX2 expressions. HUVECs were transduced with Lenti-Jag1-APEX2, exposed to static or shear conditions, and blotted for Jag1 and Actin. (D and E) Immunofluorescence microscopy of cells labeled with streptavidin 488 (SA488, green) and 4',6-diamidino-2-phylindole (DAPI; blue). MCF7 cells were transduced with (D) Lenti-APEX2-NES or (E) Lenti-Jag1-APEX2, biotinylated, and stained with streptavidin 488 and DAPI; scale bars, 25 μm.

    Article Snippet: As control plasmid for the proximity labeling we used an APEX2 domain coupled to a nuclear export signal (NES) for cytoplasmic labeling. pcDNA3 APEX2-NES (RRID:Addgene_49386) was a gift from Alice Ting through Addgene.

    Techniques: Labeling, Mass Spectrometry, Shear, Western Blot, Transduction, Immunofluorescence, Microscopy, Staining

    (A) Shown is the current model of the D. melanogaster SC. (B) Top view model of the D. melangaster SC with a schematic of the proximity ligation by APEX2::C(2)M and Cona::APEX2. APEX2::C(2)M and Cona::APEX2 incorporate into the LE and central region of the SC. In the presence of biotin-phenol and H2O2 APEX2 transfers biotin to electron-rich amino acids on nearby proteins. Nuclei are isolated from biotin-phenol treated ovaries and the biotinylated proteins were isolated on streptavidin coated beads for mass spectrometry analysis. Adapted from ( Cahoon et al . 2017 ).

    Journal: bioRxiv

    Article Title: A proximity labeling approach to identify proteins that associate with synaptonemal complex components in Drosophila melanogaster females

    doi: 10.1101/2025.10.14.682398

    Figure Lengend Snippet: (A) Shown is the current model of the D. melanogaster SC. (B) Top view model of the D. melangaster SC with a schematic of the proximity ligation by APEX2::C(2)M and Cona::APEX2. APEX2::C(2)M and Cona::APEX2 incorporate into the LE and central region of the SC. In the presence of biotin-phenol and H2O2 APEX2 transfers biotin to electron-rich amino acids on nearby proteins. Nuclei are isolated from biotin-phenol treated ovaries and the biotinylated proteins were isolated on streptavidin coated beads for mass spectrometry analysis. Adapted from ( Cahoon et al . 2017 ).

    Article Snippet: APEX2 was PCR amplified from pcDNA3 APEX2-NES (Addgene Plasmid #49386) without both the stop codon and nuclear export signal with NotI and NdeI restriction sites.

    Techniques: Ligation, Isolation, Mass Spectrometry

    Journal: bioRxiv

    Article Title: A proximity labeling approach to identify proteins that associate with synaptonemal complex components in Drosophila melanogaster females

    doi: 10.1101/2025.10.14.682398

    Figure Lengend Snippet:

    Article Snippet: APEX2 was PCR amplified from pcDNA3 APEX2-NES (Addgene Plasmid #49386) without both the stop codon and nuclear export signal with NotI and NdeI restriction sites.

    Techniques: Expressing, Construct

    Ovaries underwent biotin-phenol labeling followed by immunofluorescence with an antibody against C(3)G (SC, magenta), strevadin-488 (recognizes biotin, yellow), and DAPI (DNA, cyan). Germaria from (A) w 1118 and females expressing APEX2 did not show similar localization patterns between streptavidin-488 and C(3)G. In APEX2::c(2)M (C) and cona::APEX2 (D) germaria streptavidin localized in a pattern similar to full-length SC in a subset of nuclei. Boxed regions in the first column are shown enlarged without DAPI. Images are projections from z-stacks. Scale= 10 µm in germaria images and 2 µm in enlarged nuclei images. Full genotypes are w 1118 , y w; pUASp-APEX2/+; nanosGAL4::VP16/+; spa pol /+ , y w;; pUASp-APEX2::c(2)M; nanosGAL4::VP16/+; spa pol /+ , and y w; pUASp-cona::APEX2/+; nanosGAL4::VP16/cona f04903 ; spa pol /+ .

    Journal: bioRxiv

    Article Title: A proximity labeling approach to identify proteins that associate with synaptonemal complex components in Drosophila melanogaster females

    doi: 10.1101/2025.10.14.682398

    Figure Lengend Snippet: Ovaries underwent biotin-phenol labeling followed by immunofluorescence with an antibody against C(3)G (SC, magenta), strevadin-488 (recognizes biotin, yellow), and DAPI (DNA, cyan). Germaria from (A) w 1118 and females expressing APEX2 did not show similar localization patterns between streptavidin-488 and C(3)G. In APEX2::c(2)M (C) and cona::APEX2 (D) germaria streptavidin localized in a pattern similar to full-length SC in a subset of nuclei. Boxed regions in the first column are shown enlarged without DAPI. Images are projections from z-stacks. Scale= 10 µm in germaria images and 2 µm in enlarged nuclei images. Full genotypes are w 1118 , y w; pUASp-APEX2/+; nanosGAL4::VP16/+; spa pol /+ , y w;; pUASp-APEX2::c(2)M; nanosGAL4::VP16/+; spa pol /+ , and y w; pUASp-cona::APEX2/+; nanosGAL4::VP16/cona f04903 ; spa pol /+ .

    Article Snippet: APEX2 was PCR amplified from pcDNA3 APEX2-NES (Addgene Plasmid #49386) without both the stop codon and nuclear export signal with NotI and NdeI restriction sites.

    Techniques: Labeling, Immunofluorescence, Expressing