doxycycline inducible ptrip vectors  (Thermo Fisher)


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    Structured Review

    Thermo Fisher doxycycline inducible ptrip vectors
    Activation of SMAD signaling at low cell density and contribution of ROS signaling. ( A ) <t>Smad7</t> overexpression changes the gene expression profile of LNCaP* cells under LD_hypo conditions. Ratios of mRNA species levels measured by RT-qPCR in <t>pTRIP-mSMAD7</t> and pTRIP-control transduced cell populations cultured under LD_hypo conditions were averaged from 2 experiments using independent cell populations. Stars indicate significance of the decreased mRNA species accumulation upon murine Smad7 overexpression. ( B ) Glutathione supplementation reduces the activation of SMAD signaling at low density. Relative mRNA levels measured by RT-qPCR under LD_hypo, LD_hypo+glutathione and MD_hypo conditions were averaged from 4–5, 2 and 7–9 cell culture experiments, respectively. Stars indicate statistical significance of the difference in mRNA levels between cells cultured in LD_hypo and LD_hypo+glutathione. ( C ) Lack of additive effects between Smad7 overexpression and glutathione treatment on cloning efficiency. Cloning efficiency of pTRIP-control and pTRIP-m Smad 7 transduced cell populations were measured under the indicated cell culture conditions. Data were averaged from 2 to 5 experiments made with 2 independent control and 2 independent Smad7 transduced cell populations processed in parallel. Stars indicate significance of the differences in cloning efficiency. Data are presented as mean ± s.d.
    Doxycycline Inducible Ptrip Vectors, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 345 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/doxycycline inducible ptrip vectors/product/Thermo Fisher
    Average 88 stars, based on 345 article reviews
    Price from $9.99 to $1999.99
    doxycycline inducible ptrip vectors - by Bioz Stars, 2020-07
    88/100 stars

    Images

    1) Product Images from "SMAD signaling and redox imbalance cooperate to induce prostate cancer cell dormancy"

    Article Title: SMAD signaling and redox imbalance cooperate to induce prostate cancer cell dormancy

    Journal: Cell Cycle

    doi: 10.1080/15384101.2015.1014145

    Activation of SMAD signaling at low cell density and contribution of ROS signaling. ( A ) Smad7 overexpression changes the gene expression profile of LNCaP* cells under LD_hypo conditions. Ratios of mRNA species levels measured by RT-qPCR in pTRIP-mSMAD7 and pTRIP-control transduced cell populations cultured under LD_hypo conditions were averaged from 2 experiments using independent cell populations. Stars indicate significance of the decreased mRNA species accumulation upon murine Smad7 overexpression. ( B ) Glutathione supplementation reduces the activation of SMAD signaling at low density. Relative mRNA levels measured by RT-qPCR under LD_hypo, LD_hypo+glutathione and MD_hypo conditions were averaged from 4–5, 2 and 7–9 cell culture experiments, respectively. Stars indicate statistical significance of the difference in mRNA levels between cells cultured in LD_hypo and LD_hypo+glutathione. ( C ) Lack of additive effects between Smad7 overexpression and glutathione treatment on cloning efficiency. Cloning efficiency of pTRIP-control and pTRIP-m Smad 7 transduced cell populations were measured under the indicated cell culture conditions. Data were averaged from 2 to 5 experiments made with 2 independent control and 2 independent Smad7 transduced cell populations processed in parallel. Stars indicate significance of the differences in cloning efficiency. Data are presented as mean ± s.d.
    Figure Legend Snippet: Activation of SMAD signaling at low cell density and contribution of ROS signaling. ( A ) Smad7 overexpression changes the gene expression profile of LNCaP* cells under LD_hypo conditions. Ratios of mRNA species levels measured by RT-qPCR in pTRIP-mSMAD7 and pTRIP-control transduced cell populations cultured under LD_hypo conditions were averaged from 2 experiments using independent cell populations. Stars indicate significance of the decreased mRNA species accumulation upon murine Smad7 overexpression. ( B ) Glutathione supplementation reduces the activation of SMAD signaling at low density. Relative mRNA levels measured by RT-qPCR under LD_hypo, LD_hypo+glutathione and MD_hypo conditions were averaged from 4–5, 2 and 7–9 cell culture experiments, respectively. Stars indicate statistical significance of the difference in mRNA levels between cells cultured in LD_hypo and LD_hypo+glutathione. ( C ) Lack of additive effects between Smad7 overexpression and glutathione treatment on cloning efficiency. Cloning efficiency of pTRIP-control and pTRIP-m Smad 7 transduced cell populations were measured under the indicated cell culture conditions. Data were averaged from 2 to 5 experiments made with 2 independent control and 2 independent Smad7 transduced cell populations processed in parallel. Stars indicate significance of the differences in cloning efficiency. Data are presented as mean ± s.d.

    Techniques Used: Activation Assay, Over Expression, Expressing, Quantitative RT-PCR, Cell Culture, Clone Assay

    2) Product Images from "SMAD signaling and redox imbalance cooperate to induce prostate cancer cell dormancy"

    Article Title: SMAD signaling and redox imbalance cooperate to induce prostate cancer cell dormancy

    Journal: Cell Cycle

    doi: 10.1080/15384101.2015.1014145

    Activation of SMAD signaling at low cell density and contribution of ROS signaling. ( A ) Smad7 overexpression changes the gene expression profile of LNCaP* cells under LD_hypo conditions. Ratios of mRNA species levels measured by RT-qPCR in pTRIP-mSMAD7 and pTRIP-control transduced cell populations cultured under LD_hypo conditions were averaged from 2 experiments using independent cell populations. Stars indicate significance of the decreased mRNA species accumulation upon murine Smad7 overexpression. ( B ) Glutathione supplementation reduces the activation of SMAD signaling at low density. Relative mRNA levels measured by RT-qPCR under LD_hypo, LD_hypo+glutathione and MD_hypo conditions were averaged from 4–5, 2 and 7–9 cell culture experiments, respectively. Stars indicate statistical significance of the difference in mRNA levels between cells cultured in LD_hypo and LD_hypo+glutathione. ( C ) Lack of additive effects between Smad7 overexpression and glutathione treatment on cloning efficiency. Cloning efficiency of pTRIP-control and pTRIP-m Smad 7 transduced cell populations were measured under the indicated cell culture conditions. Data were averaged from 2 to 5 experiments made with 2 independent control and 2 independent Smad7 transduced cell populations processed in parallel. Stars indicate significance of the differences in cloning efficiency. Data are presented as mean ± s.d.
    Figure Legend Snippet: Activation of SMAD signaling at low cell density and contribution of ROS signaling. ( A ) Smad7 overexpression changes the gene expression profile of LNCaP* cells under LD_hypo conditions. Ratios of mRNA species levels measured by RT-qPCR in pTRIP-mSMAD7 and pTRIP-control transduced cell populations cultured under LD_hypo conditions were averaged from 2 experiments using independent cell populations. Stars indicate significance of the decreased mRNA species accumulation upon murine Smad7 overexpression. ( B ) Glutathione supplementation reduces the activation of SMAD signaling at low density. Relative mRNA levels measured by RT-qPCR under LD_hypo, LD_hypo+glutathione and MD_hypo conditions were averaged from 4–5, 2 and 7–9 cell culture experiments, respectively. Stars indicate statistical significance of the difference in mRNA levels between cells cultured in LD_hypo and LD_hypo+glutathione. ( C ) Lack of additive effects between Smad7 overexpression and glutathione treatment on cloning efficiency. Cloning efficiency of pTRIP-control and pTRIP-m Smad 7 transduced cell populations were measured under the indicated cell culture conditions. Data were averaged from 2 to 5 experiments made with 2 independent control and 2 independent Smad7 transduced cell populations processed in parallel. Stars indicate significance of the differences in cloning efficiency. Data are presented as mean ± s.d.

    Techniques Used: Activation Assay, Over Expression, Expressing, Quantitative RT-PCR, Cell Culture, Clone Assay

    3) Product Images from "p97-dependent retrotranslocation and proteolytic processing govern formation of active Nrf1 upon proteasome inhibition"

    Article Title: p97-dependent retrotranslocation and proteolytic processing govern formation of active Nrf1 upon proteasome inhibition

    Journal: eLife

    doi: 10.7554/eLife.01856

    p97 is required for processing and activation of Nrf1. ( A ) NIH-3T3-p97sh cells stably expressing doxycycline (Doxy)-inducible shRNA targeting p97 were either mock-treated or induced with Doxy (1 µg/ml) for 3 days after which the cells were further treated with DMSO or 1 µM MG132 as indicated for 10 hr. The RNA from these cells was used for quantitative RT-PCR to assess the mRNA levels of the representative PSM genes. The values were normalized to GAPDH mRNA levels. Error bars denote SD (n = 3). ( B ) NIH-3T3-p97sh cells were subjected to Doxy and MG132 treatments as above and the cell lysates were fractionated by SDS-PAGE and immunoblotted to detect p97 and Nrf1. β-actin was used as a loading control. ( C ) NIH-3T3 (left panel) or HEK-293-Nrf1 3×Flag cells (right panel) were pulsed for an hour with 10 µM NMS-873 and then chased with MG132 plus cycloheximide (CHX). The cells were harvested at the time points indicated (from 0 min to 120 min) and the lysates were fractionated by SDS-PAGE and immunoblotted to detect endogenous Nrf1. β-actin was used as a loading control. DOI: http://dx.doi.org/10.7554/eLife.01856.005
    Figure Legend Snippet: p97 is required for processing and activation of Nrf1. ( A ) NIH-3T3-p97sh cells stably expressing doxycycline (Doxy)-inducible shRNA targeting p97 were either mock-treated or induced with Doxy (1 µg/ml) for 3 days after which the cells were further treated with DMSO or 1 µM MG132 as indicated for 10 hr. The RNA from these cells was used for quantitative RT-PCR to assess the mRNA levels of the representative PSM genes. The values were normalized to GAPDH mRNA levels. Error bars denote SD (n = 3). ( B ) NIH-3T3-p97sh cells were subjected to Doxy and MG132 treatments as above and the cell lysates were fractionated by SDS-PAGE and immunoblotted to detect p97 and Nrf1. β-actin was used as a loading control. ( C ) NIH-3T3 (left panel) or HEK-293-Nrf1 3×Flag cells (right panel) were pulsed for an hour with 10 µM NMS-873 and then chased with MG132 plus cycloheximide (CHX). The cells were harvested at the time points indicated (from 0 min to 120 min) and the lysates were fractionated by SDS-PAGE and immunoblotted to detect endogenous Nrf1. β-actin was used as a loading control. DOI: http://dx.doi.org/10.7554/eLife.01856.005

    Techniques Used: Activation Assay, Stable Transfection, Expressing, shRNA, Quantitative RT-PCR, SDS Page

    Related Articles

    Clone Assay:

    Article Title: Switching and emergence of CTL epitopes in HIV-1 infection
    Article Snippet: .. CTL clones were cultured with RPMI 1640 supplemented with 50 U of interleukin-2/ml, 100U of penicillin/ml, 100U of streptomycin/ml, and 10% heat-inactivated FCS (R10/50), but the clones were cultured in the absence of interleukin-2 (R10) for two days before antigen presentation assays. pcDNA3.1-A24-DsRedm was introduced into 293FT cell line (Invitrogen) and the cells were treated by hygromycin for 2 weeks. .. After cloning by limiting dilution we obtained 293FT-A24DRm-CY0, and confirmed HLA-A*24:02 expression with FACS analysis by using anti-HLA-A9 serotype antibody (One Lambda, data not shown).

    Transfection:

    Article Title: RasGRP3 Mediates MAPK Pathway Activation in GNAQ Mutant Uveal Melanoma
    Article Snippet: .. For transient transfection, 293FT cells were transfected using lipofectamine 2000(Invitrogen) following the manufacturer’s instruction. .. Lentiviral generation and transduction have been described previou sly ( ).

    Article Title: Design and Characterization of a Computationally Optimized Broadly Reactive Hemagglutinin Vaccine for H1N1 Influenza Viruses
    Article Snippet: .. To confirm antigenicity of cell surface-expressed COBRA HAs, HEK 293FT cells (1 × 106 ) were transiently transfected with 2 μg DNA encoding each COBRA HA or wild-type HA using Lipofectamine 2000 (Life Technologies) according to the manufacturer's instructions. .. Transfected cells were harvested 24 h posttransfection by gentle dissociation with TrypLE Express (Life Technologies) and labeled by using a LIVE/DEAD Fixable Far Red Dead Cell Stain kit (Life Technologies) to determine cell viability.

    Article Title: Androgen receptor serine 81 mediates Pin1 interaction and activity
    Article Snippet: .. Briefly, 1 × 106 293FT cells (Invitrogen Corp.) were transfected with 2.25 μg of PAX2 packaging plasmid, 0.75 μg of PMD2G envelope plasmid and 3 μg of pLKO.1 hairpin vector utilizing 30 μl of Fugene HD (Roche Applied Science) in 10 cm plates. .. Polyclonal populations of Pin1 kd or overexpressed and scrambled cells were generated by infection with 1 MOI (multiplicity of infectious) of lentiviral particles.

    Article Title: Transfection of shRNA-encoding Minivector DNA of a few hundred base pairs to regulate gene expression in lymphoma cells
    Article Snippet: .. Cell transfection/gene silencing As a reporter system for GFP gene silencing, we established stable GFP -expressing cells from adhesion 293FT cells (a transformed human embryonic kidney cell line, Invitrogen) or suspension Jurkat cells (a human T-lymphoma/leukemia cell line from ATCC, Manassas, VA, USA). .. The GFP -expressing cells were transfected with 1 μg of Minivector DNA or plasmid (given the difference between the sizes, ∼10-fold more moles of Minivector are delivered relative to plasmid) or 20 pmol synthetic siRNAs using Lipofectamine methodology following the manufacturer's instructions (Invitrogen).

    Article Title: Characterization of a Merkel Cell Polyomavirus-Positive Merkel Cell Carcinoma Cell Line CVG-1
    Article Snippet: .. For lentivirus production, 293FT cells (Invitrogen) were transfected with pLenti TRE MCV sT, psPax2, and pMD2.G using Lipofectamine 2000 (Invitrogen). .. The medium was changed at 12 h after transfection, and the lentivirus produced in the culture supernatant was harvested at 72 h after transfection.

    Combined Bisulfite Restriction Analysis Assay:

    Article Title: Design and Characterization of a Computationally Optimized Broadly Reactive Hemagglutinin Vaccine for H1N1 Influenza Viruses
    Article Snippet: .. To confirm antigenicity of cell surface-expressed COBRA HAs, HEK 293FT cells (1 × 106 ) were transiently transfected with 2 μg DNA encoding each COBRA HA or wild-type HA using Lipofectamine 2000 (Life Technologies) according to the manufacturer's instructions. .. Transfected cells were harvested 24 h posttransfection by gentle dissociation with TrypLE Express (Life Technologies) and labeled by using a LIVE/DEAD Fixable Far Red Dead Cell Stain kit (Life Technologies) to determine cell viability.

    Cell Culture:

    Article Title: The DNA Damage Sensors Ataxia-Telangiectasia Mutated Kinase and Checkpoint Kinase 2 Are Required for Hepatitis C Virus RNA Replication ▿
    Article Snippet: .. 293FT cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS). .. The HuH-7-derived O cells harboring a replicative genome-length HCV RNA and the HuH-7-derived sO cells harboring the subgenomic replicon RNA of HCV-O were cultured in DMEM with 10% FBS and G418 (300 μg/ml geneticin; Invitrogen) as described previously ( , ).

    Article Title: Switching and emergence of CTL epitopes in HIV-1 infection
    Article Snippet: .. CTL clones were cultured with RPMI 1640 supplemented with 50 U of interleukin-2/ml, 100U of penicillin/ml, 100U of streptomycin/ml, and 10% heat-inactivated FCS (R10/50), but the clones were cultured in the absence of interleukin-2 (R10) for two days before antigen presentation assays. pcDNA3.1-A24-DsRedm was introduced into 293FT cell line (Invitrogen) and the cells were treated by hygromycin for 2 weeks. .. After cloning by limiting dilution we obtained 293FT-A24DRm-CY0, and confirmed HLA-A*24:02 expression with FACS analysis by using anti-HLA-A9 serotype antibody (One Lambda, data not shown).

    CTL Assay:

    Article Title: Switching and emergence of CTL epitopes in HIV-1 infection
    Article Snippet: .. CTL clones were cultured with RPMI 1640 supplemented with 50 U of interleukin-2/ml, 100U of penicillin/ml, 100U of streptomycin/ml, and 10% heat-inactivated FCS (R10/50), but the clones were cultured in the absence of interleukin-2 (R10) for two days before antigen presentation assays. pcDNA3.1-A24-DsRedm was introduced into 293FT cell line (Invitrogen) and the cells were treated by hygromycin for 2 weeks. .. After cloning by limiting dilution we obtained 293FT-A24DRm-CY0, and confirmed HLA-A*24:02 expression with FACS analysis by using anti-HLA-A9 serotype antibody (One Lambda, data not shown).

    Expressing:

    Article Title: Transfection of shRNA-encoding Minivector DNA of a few hundred base pairs to regulate gene expression in lymphoma cells
    Article Snippet: .. Cell transfection/gene silencing As a reporter system for GFP gene silencing, we established stable GFP -expressing cells from adhesion 293FT cells (a transformed human embryonic kidney cell line, Invitrogen) or suspension Jurkat cells (a human T-lymphoma/leukemia cell line from ATCC, Manassas, VA, USA). .. The GFP -expressing cells were transfected with 1 μg of Minivector DNA or plasmid (given the difference between the sizes, ∼10-fold more moles of Minivector are delivered relative to plasmid) or 20 pmol synthetic siRNAs using Lipofectamine methodology following the manufacturer's instructions (Invitrogen).

    Modification:

    Article Title: The DNA Damage Sensors Ataxia-Telangiectasia Mutated Kinase and Checkpoint Kinase 2 Are Required for Hepatitis C Virus RNA Replication ▿
    Article Snippet: .. 293FT cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS). .. The HuH-7-derived O cells harboring a replicative genome-length HCV RNA and the HuH-7-derived sO cells harboring the subgenomic replicon RNA of HCV-O were cultured in DMEM with 10% FBS and G418 (300 μg/ml geneticin; Invitrogen) as described previously ( , ).

    Article Title: Biological effects of CCS in the absence of SOD1 enzyme activation: implications for disease in a mouse model for ALS
    Article Snippet: .. HEK 293 and HEK 293FT cells were maintained in high-glucose Dulbecco's modified Eagle's medium supplemented with either 10% fetal bovine serum/1 mM L-glutamine or 10% fetal bovine serum/2 m m L-glutamine/1 m m L-pyruvate/1:100 MEM non-essential amino acids (Invitrogen)/500 ug/ml Geneticin respectively, using standard culture procedures. .. Transient transfections in HEK 293 and HEK 293FT were performed on > 90% confluent cells in 60 mm dishes (poly-d-lysine coated for FT cells; BD Biocoat) using Lipofectamine 2000 transfection reagent (Invitrogen) (10 µl/500 µl for a 60 mm plate).

    Transformation Assay:

    Article Title: Transfection of shRNA-encoding Minivector DNA of a few hundred base pairs to regulate gene expression in lymphoma cells
    Article Snippet: .. Cell transfection/gene silencing As a reporter system for GFP gene silencing, we established stable GFP -expressing cells from adhesion 293FT cells (a transformed human embryonic kidney cell line, Invitrogen) or suspension Jurkat cells (a human T-lymphoma/leukemia cell line from ATCC, Manassas, VA, USA). .. The GFP -expressing cells were transfected with 1 μg of Minivector DNA or plasmid (given the difference between the sizes, ∼10-fold more moles of Minivector are delivered relative to plasmid) or 20 pmol synthetic siRNAs using Lipofectamine methodology following the manufacturer's instructions (Invitrogen).

    Plasmid Preparation:

    Article Title: Androgen receptor serine 81 mediates Pin1 interaction and activity
    Article Snippet: .. Briefly, 1 × 106 293FT cells (Invitrogen Corp.) were transfected with 2.25 μg of PAX2 packaging plasmid, 0.75 μg of PMD2G envelope plasmid and 3 μg of pLKO.1 hairpin vector utilizing 30 μl of Fugene HD (Roche Applied Science) in 10 cm plates. .. Polyclonal populations of Pin1 kd or overexpressed and scrambled cells were generated by infection with 1 MOI (multiplicity of infectious) of lentiviral particles.

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