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Stratagene plasmid pcmv lacz
Plasmid Pcmv Lacz, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/plasmid pcmv lacz/product/Stratagene
Average 85 stars, based on 1 article reviews
Price from $9.99 to $1999.99
plasmid pcmv lacz - by Bioz Stars, 2020-09
85/100 stars

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Article Title: Involvement of the Rac1-IRSp53-Wave2-Arp2/3 Signaling Pathway in HIV-1 Gag Particle Release in CD4 T Cells
Article Snippet: .. Plasmids pCMV-LacZ and pCMV-GFP were used as control plasmids and were derived from a pCMV tag plasmid (Stratagene, Agilent Technologies, Madrid, Spain) that expresses green fluorescent protein (GFP) or LacZ under the control of the cytomegalovirus (CMV) promoter. .. Jurkat T cells (2 × 106 ) were microporated with 5 μg of p8.2 or pNL4.3ΔEnv or 4 μg of pGag, pCMV-LacZ, or pCMV-GFP (as control plasmids), together with 150 pmol Stealth RNA interference (RNAi) siRNA against Rho GTPases (Invitrogen); 360 pmol Stealth RNAi siRNA against the Rho GTPase effector Wave2, Vav1, Dia1, or Pak2 (Invitrogen); or 250 pmol siRNA against IRSp53 or Arp3 (Santa Cruz) or the corresponding siRNA controls.

Plasmid Preparation:

Article Title: Involvement of the Rac1-IRSp53-Wave2-Arp2/3 Signaling Pathway in HIV-1 Gag Particle Release in CD4 T Cells
Article Snippet: .. Plasmids pCMV-LacZ and pCMV-GFP were used as control plasmids and were derived from a pCMV tag plasmid (Stratagene, Agilent Technologies, Madrid, Spain) that expresses green fluorescent protein (GFP) or LacZ under the control of the cytomegalovirus (CMV) promoter. .. Jurkat T cells (2 × 106 ) were microporated with 5 μg of p8.2 or pNL4.3ΔEnv or 4 μg of pGag, pCMV-LacZ, or pCMV-GFP (as control plasmids), together with 150 pmol Stealth RNA interference (RNAi) siRNA against Rho GTPases (Invitrogen); 360 pmol Stealth RNAi siRNA against the Rho GTPase effector Wave2, Vav1, Dia1, or Pak2 (Invitrogen); or 250 pmol siRNA against IRSp53 or Arp3 (Santa Cruz) or the corresponding siRNA controls.

Article Title: Activation of c-myc promoter P1 by immunoglobulin ? gene enhancers in Burkitt lymphoma: functional characterization of the intron enhancer motifs ?B, E box 1 and E box 2, and of the 3? enhancer motif PU
Article Snippet: .. Plasmid pCMV-lacZ was obtained from Stratagene (USA). ..

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    Stratagene plasmids pcmv lacz
    Effect of a partial double knockdown of IRSp53/Wave2 by siRNA on HIV-1 Gag localization and VLP release in T cells. (A) Jurkat T cells were transfected with the vector pGag (HIV-1 Gag expression) (a to d) or <t>pCMV-LacZ</t> (control) (e and f), together with a mix of siRNA (Wave2 and IRSp53) (a and f) or a mix of control siRNAs (d). The cells were fixed and stained for F-actin (phalloidin-Alexa 546) (red) and Gag (anti-MAp17-Alexa 488) (green). Merge and transmission images are presented. The cell sections show Gag and F-actin fluorescent signals at the cell periphery. (B) Distance of the maximum Gag fluorescent signal from the cell edge (determined by the actin signal) under each condition ( n = 9 to 21 cells). (C) Cell mean diameters (in micrometers) of the different cell transfection conditions, as indicated. The effect of the depletion of IRSp53 and/or Wave2 on Jurkat T-cell sizes compared to those of siRNA control-treated cells, in the presence of Gag, is shown. (D) Immunoblot analysis of pr55Gag in T-cell lysates and in VLPs. On the right are the average percentages of depletion of each protein and of inhibition of VLP release from three independent experiments. Tubulin was used as a loading control.
    Plasmids Pcmv Lacz, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmids pcmv lacz/product/Stratagene
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    plasmids pcmv lacz - by Bioz Stars, 2020-09
    88/100 stars
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    Effect of a partial double knockdown of IRSp53/Wave2 by siRNA on HIV-1 Gag localization and VLP release in T cells. (A) Jurkat T cells were transfected with the vector pGag (HIV-1 Gag expression) (a to d) or pCMV-LacZ (control) (e and f), together with a mix of siRNA (Wave2 and IRSp53) (a and f) or a mix of control siRNAs (d). The cells were fixed and stained for F-actin (phalloidin-Alexa 546) (red) and Gag (anti-MAp17-Alexa 488) (green). Merge and transmission images are presented. The cell sections show Gag and F-actin fluorescent signals at the cell periphery. (B) Distance of the maximum Gag fluorescent signal from the cell edge (determined by the actin signal) under each condition ( n = 9 to 21 cells). (C) Cell mean diameters (in micrometers) of the different cell transfection conditions, as indicated. The effect of the depletion of IRSp53 and/or Wave2 on Jurkat T-cell sizes compared to those of siRNA control-treated cells, in the presence of Gag, is shown. (D) Immunoblot analysis of pr55Gag in T-cell lysates and in VLPs. On the right are the average percentages of depletion of each protein and of inhibition of VLP release from three independent experiments. Tubulin was used as a loading control.

    Journal: Journal of Virology

    Article Title: Involvement of the Rac1-IRSp53-Wave2-Arp2/3 Signaling Pathway in HIV-1 Gag Particle Release in CD4 T Cells

    doi: 10.1128/JVI.00469-15

    Figure Lengend Snippet: Effect of a partial double knockdown of IRSp53/Wave2 by siRNA on HIV-1 Gag localization and VLP release in T cells. (A) Jurkat T cells were transfected with the vector pGag (HIV-1 Gag expression) (a to d) or pCMV-LacZ (control) (e and f), together with a mix of siRNA (Wave2 and IRSp53) (a and f) or a mix of control siRNAs (d). The cells were fixed and stained for F-actin (phalloidin-Alexa 546) (red) and Gag (anti-MAp17-Alexa 488) (green). Merge and transmission images are presented. The cell sections show Gag and F-actin fluorescent signals at the cell periphery. (B) Distance of the maximum Gag fluorescent signal from the cell edge (determined by the actin signal) under each condition ( n = 9 to 21 cells). (C) Cell mean diameters (in micrometers) of the different cell transfection conditions, as indicated. The effect of the depletion of IRSp53 and/or Wave2 on Jurkat T-cell sizes compared to those of siRNA control-treated cells, in the presence of Gag, is shown. (D) Immunoblot analysis of pr55Gag in T-cell lysates and in VLPs. On the right are the average percentages of depletion of each protein and of inhibition of VLP release from three independent experiments. Tubulin was used as a loading control.

    Article Snippet: Plasmids pCMV-LacZ and pCMV-GFP were used as control plasmids and were derived from a pCMV tag plasmid (Stratagene, Agilent Technologies, Madrid, Spain) that expresses green fluorescent protein (GFP) or LacZ under the control of the cytomegalovirus (CMV) promoter.

    Techniques: Transfection, Plasmid Preparation, Expressing, Staining, Transmission Assay, Inhibition

    Effect of siRNA-mediated depletion of Rac1 on intracellular PI(4,5)P 2 levels in Jurkat T cells. (A) Rac1-dependent cell signaling pathways having an effect on actin filament dynamics. In T cells, Rac1 GTPase plays an important role in the regulation of actin cytoskeleton rearrangement. Indeed, on one hand, in response to extracellular signals, Rac1 is activated and could stimulate actin filament turnover by the intermediate of the Pak1 (or Pak2)-LIMK-cofilin pathway. On the other hand, activated Rac1 could also stimulate actin filament branching in lamellipodia. In the latter case, IRSp53 is recruited by Rac1 and binds the proline-rich region of Wave2. As a result, Wave2 is activated and induces actin branching via Arp2/3 complex recruitment. In a third case, Rac1-GTP could also activate the phosphatidylinositol-4-phosphate 5-kinase (PIP5K), which catalyzes the synthesis of PI(4,5)P 2 at the cell plasma membrane. (B to D) Effect of siRNA-mediated depletion of Rac1 on intracellular PI(4,5)P 2 levels. (B) Cells were transfected with pCMV-LacZ, p8.2 (Gag, Gag-Pol, and viral accessory proteins), or PH-phospholipase C delta (PLCδ)-GFP, together with the siRNA control or siRNA against Rac1, as indicated. At 48 h posttransfection, cells were fixed, permeabilized, and stained with anti-MAp17 and anti-PI(4,5)P 2 antibodies. The colocalization of the GFP-tagged PH domain of phospholipase C delta and the PI(4,5)P2 antibody is indicated as a control for PI(4,5)P2-enriched membrane domains (white arrows). (C) The ratio of the PI(4,5)P 2 signal intensity to the cell area was measured by image analysis (ImageJ) ( n = 20 cells). (D) Immunoblot analysis of Rac1 GTPase and HIV-1 Gag in cell lysates and/or in virus particles. Tubulin was used as a loading control.

    Journal: Journal of Virology

    Article Title: Involvement of the Rac1-IRSp53-Wave2-Arp2/3 Signaling Pathway in HIV-1 Gag Particle Release in CD4 T Cells

    doi: 10.1128/JVI.00469-15

    Figure Lengend Snippet: Effect of siRNA-mediated depletion of Rac1 on intracellular PI(4,5)P 2 levels in Jurkat T cells. (A) Rac1-dependent cell signaling pathways having an effect on actin filament dynamics. In T cells, Rac1 GTPase plays an important role in the regulation of actin cytoskeleton rearrangement. Indeed, on one hand, in response to extracellular signals, Rac1 is activated and could stimulate actin filament turnover by the intermediate of the Pak1 (or Pak2)-LIMK-cofilin pathway. On the other hand, activated Rac1 could also stimulate actin filament branching in lamellipodia. In the latter case, IRSp53 is recruited by Rac1 and binds the proline-rich region of Wave2. As a result, Wave2 is activated and induces actin branching via Arp2/3 complex recruitment. In a third case, Rac1-GTP could also activate the phosphatidylinositol-4-phosphate 5-kinase (PIP5K), which catalyzes the synthesis of PI(4,5)P 2 at the cell plasma membrane. (B to D) Effect of siRNA-mediated depletion of Rac1 on intracellular PI(4,5)P 2 levels. (B) Cells were transfected with pCMV-LacZ, p8.2 (Gag, Gag-Pol, and viral accessory proteins), or PH-phospholipase C delta (PLCδ)-GFP, together with the siRNA control or siRNA against Rac1, as indicated. At 48 h posttransfection, cells were fixed, permeabilized, and stained with anti-MAp17 and anti-PI(4,5)P 2 antibodies. The colocalization of the GFP-tagged PH domain of phospholipase C delta and the PI(4,5)P2 antibody is indicated as a control for PI(4,5)P2-enriched membrane domains (white arrows). (C) The ratio of the PI(4,5)P 2 signal intensity to the cell area was measured by image analysis (ImageJ) ( n = 20 cells). (D) Immunoblot analysis of Rac1 GTPase and HIV-1 Gag in cell lysates and/or in virus particles. Tubulin was used as a loading control.

    Article Snippet: Plasmids pCMV-LacZ and pCMV-GFP were used as control plasmids and were derived from a pCMV tag plasmid (Stratagene, Agilent Technologies, Madrid, Spain) that expresses green fluorescent protein (GFP) or LacZ under the control of the cytomegalovirus (CMV) promoter.

    Techniques: Transfection, Staining