retro x tet on inducible cell lines  (TaKaRa)


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    TaKaRa retro x tet on inducible cell lines
    The ssDNA binding ability of SDE2 is required for the function of the fork protection complex (FPC) at replication forks (A) Top: <t>Retro-X</t> <t>Tet-One</t> system. Doxycycline (dox) induces the expression of siRNA-resistant cDNA under the P TRE3GS promoter upon its binding to the <t>Tet-On</t> 3G transactivator. Bottom: induction of SDE2 WT or ΔSAP (Δ395-451) in response to dox, following siRNA transfection. (B) Subcellular fractionation of U2OS <t>cells</t> re-expressing SDE2 WT or ΔSAP into S (cytosolic), P1 (nuclear, non-chromatin), and P2 (chromatin) fractions. (C) Representative images of TIM:EdU PLA foci in the Retro-X SDE2 WT or ΔSAP cells following SDE2 siRNA transfection and dox induction. Scale bar: 10 μm. (D) Left: quantification of cells positive for TIM-EdU PLA foci ( > 400 cells per condition, n=3, error=SEM, ** p
    Retro X Tet On Inducible Cell Lines, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/retro x tet on inducible cell lines/product/TaKaRa
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    retro x tet on inducible cell lines - by Bioz Stars, 2022-09
    94/100 stars

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    1) Product Images from "Extended DNA binding interface beyond the canonical SAP domain contributes to SDE2 function at DNA replication forks"

    Article Title: Extended DNA binding interface beyond the canonical SAP domain contributes to SDE2 function at DNA replication forks

    Journal: bioRxiv

    doi: 10.1101/2022.05.09.490802

    The ssDNA binding ability of SDE2 is required for the function of the fork protection complex (FPC) at replication forks (A) Top: Retro-X Tet-One system. Doxycycline (dox) induces the expression of siRNA-resistant cDNA under the P TRE3GS promoter upon its binding to the Tet-On 3G transactivator. Bottom: induction of SDE2 WT or ΔSAP (Δ395-451) in response to dox, following siRNA transfection. (B) Subcellular fractionation of U2OS cells re-expressing SDE2 WT or ΔSAP into S (cytosolic), P1 (nuclear, non-chromatin), and P2 (chromatin) fractions. (C) Representative images of TIM:EdU PLA foci in the Retro-X SDE2 WT or ΔSAP cells following SDE2 siRNA transfection and dox induction. Scale bar: 10 μm. (D) Left: quantification of cells positive for TIM-EdU PLA foci ( > 400 cells per condition, n=3, error=SEM, ** p
    Figure Legend Snippet: The ssDNA binding ability of SDE2 is required for the function of the fork protection complex (FPC) at replication forks (A) Top: Retro-X Tet-One system. Doxycycline (dox) induces the expression of siRNA-resistant cDNA under the P TRE3GS promoter upon its binding to the Tet-On 3G transactivator. Bottom: induction of SDE2 WT or ΔSAP (Δ395-451) in response to dox, following siRNA transfection. (B) Subcellular fractionation of U2OS cells re-expressing SDE2 WT or ΔSAP into S (cytosolic), P1 (nuclear, non-chromatin), and P2 (chromatin) fractions. (C) Representative images of TIM:EdU PLA foci in the Retro-X SDE2 WT or ΔSAP cells following SDE2 siRNA transfection and dox induction. Scale bar: 10 μm. (D) Left: quantification of cells positive for TIM-EdU PLA foci ( > 400 cells per condition, n=3, error=SEM, ** p

    Techniques Used: Binding Assay, Expressing, Transfection, Fractionation, Proximity Ligation Assay

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  • 94
    TaKaRa retro x tet on inducible cell lines
    The ssDNA binding ability of SDE2 is required for the function of the fork protection complex (FPC) at replication forks (A) Top: <t>Retro-X</t> <t>Tet-One</t> system. Doxycycline (dox) induces the expression of siRNA-resistant cDNA under the P TRE3GS promoter upon its binding to the <t>Tet-On</t> 3G transactivator. Bottom: induction of SDE2 WT or ΔSAP (Δ395-451) in response to dox, following siRNA transfection. (B) Subcellular fractionation of U2OS <t>cells</t> re-expressing SDE2 WT or ΔSAP into S (cytosolic), P1 (nuclear, non-chromatin), and P2 (chromatin) fractions. (C) Representative images of TIM:EdU PLA foci in the Retro-X SDE2 WT or ΔSAP cells following SDE2 siRNA transfection and dox induction. Scale bar: 10 μm. (D) Left: quantification of cells positive for TIM-EdU PLA foci ( > 400 cells per condition, n=3, error=SEM, ** p
    Retro X Tet On Inducible Cell Lines, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/retro x tet on inducible cell lines/product/TaKaRa
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    retro x tet on inducible cell lines - by Bioz Stars, 2022-09
    94/100 stars
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    The ssDNA binding ability of SDE2 is required for the function of the fork protection complex (FPC) at replication forks (A) Top: Retro-X Tet-One system. Doxycycline (dox) induces the expression of siRNA-resistant cDNA under the P TRE3GS promoter upon its binding to the Tet-On 3G transactivator. Bottom: induction of SDE2 WT or ΔSAP (Δ395-451) in response to dox, following siRNA transfection. (B) Subcellular fractionation of U2OS cells re-expressing SDE2 WT or ΔSAP into S (cytosolic), P1 (nuclear, non-chromatin), and P2 (chromatin) fractions. (C) Representative images of TIM:EdU PLA foci in the Retro-X SDE2 WT or ΔSAP cells following SDE2 siRNA transfection and dox induction. Scale bar: 10 μm. (D) Left: quantification of cells positive for TIM-EdU PLA foci ( > 400 cells per condition, n=3, error=SEM, ** p

    Journal: bioRxiv

    Article Title: Extended DNA binding interface beyond the canonical SAP domain contributes to SDE2 function at DNA replication forks

    doi: 10.1101/2022.05.09.490802

    Figure Lengend Snippet: The ssDNA binding ability of SDE2 is required for the function of the fork protection complex (FPC) at replication forks (A) Top: Retro-X Tet-One system. Doxycycline (dox) induces the expression of siRNA-resistant cDNA under the P TRE3GS promoter upon its binding to the Tet-On 3G transactivator. Bottom: induction of SDE2 WT or ΔSAP (Δ395-451) in response to dox, following siRNA transfection. (B) Subcellular fractionation of U2OS cells re-expressing SDE2 WT or ΔSAP into S (cytosolic), P1 (nuclear, non-chromatin), and P2 (chromatin) fractions. (C) Representative images of TIM:EdU PLA foci in the Retro-X SDE2 WT or ΔSAP cells following SDE2 siRNA transfection and dox induction. Scale bar: 10 μm. (D) Left: quantification of cells positive for TIM-EdU PLA foci ( > 400 cells per condition, n=3, error=SEM, ** p

    Article Snippet: Generation of Retro-X Tet-On inducible cell lines The retroviral plasmid pRetroX-TetOne puro was acquired from Clontech and amplified using NEB Stable competent E. coli (high efficiency). siRNA-1 resistant SDE2 WT or ΔSAP (deletion of amino acids 395-451 along with ΔUBL 1-77) were subcloned into pRetroX-TetOne puro vector.

    Techniques: Binding Assay, Expressing, Transfection, Fractionation, Proximity Ligation Assay

    SHP 2 catalytic activity is required for PDGF ‐evoked ERK MAPK signaling, but not for PDGFR β phosphorylation. Ptpn11 fl/fl MEF s expressing CRE ‐ ER T am with or without retroviral expression of WT or C459E mutant SHP 2 were treated with 4‐ OHT (1 μ m ) for 4 days or left untreated. Cells were then serum‐starved, followed by treatment with PDGF ‐ BB (50 ng·mL −1 ) for the indicated times. Lysates were subjected to immunoblotting with the indicated antibodies. Representative immunoblots are shown from one of three experiments.

    Journal: FEBS Open Bio

    Article Title: Off‐target inhibition by active site‐targeting SHP2 inhibitors

    doi: 10.1002/2211-5463.12493

    Figure Lengend Snippet: SHP 2 catalytic activity is required for PDGF ‐evoked ERK MAPK signaling, but not for PDGFR β phosphorylation. Ptpn11 fl/fl MEF s expressing CRE ‐ ER T am with or without retroviral expression of WT or C459E mutant SHP 2 were treated with 4‐ OHT (1 μ m ) for 4 days or left untreated. Cells were then serum‐starved, followed by treatment with PDGF ‐ BB (50 ng·mL −1 ) for the indicated times. Lysates were subjected to immunoblotting with the indicated antibodies. Representative immunoblots are shown from one of three experiments.

    Article Snippet: Expression constructs, infection, and sorting Retroviral expression vectors for wild‐type SHP2 (WT SHP2) and the mutant SHP2C459E were generated by subcloning human PTPN11 cDNA into pMSCV‐IRES‐EGFP (Clontech).

    Techniques: Activity Assay, Expressing, Mutagenesis, Western Blot