transient transfection  (Millipore)


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    Name:
    Escort IV Transfection Reagent
    Description:
    Escort TM IV is a unique formulation of a proprietary polycationic lipid and a neutral non transfecting lipid This liposome forming compound is used for transfection of nucleic acids into a wide variety of eukaryotic cell types
    Catalog Number:
    l3287
    Price:
    None
    Applications:
    Suitable for transient and stable transfection of nucleic acids into cultured eukaryotic cells. Use approximately 4-16 muL Escort(TM) IV and 2 mug DNA per 6 cm cell culture plate. Protocol optimization provides very efficient transfection. For a list of cells that have been successfully transfected using Escort(TM) IV, see the Transfection Reagent Selection Guide.
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    Structured Review

    Millipore transient transfection
    Escort IV Transfection Reagent
    Escort TM IV is a unique formulation of a proprietary polycationic lipid and a neutral non transfecting lipid This liposome forming compound is used for transfection of nucleic acids into a wide variety of eukaryotic cell types
    https://www.bioz.com/result/transient transfection/product/Millipore
    Average 99 stars, based on 1544 article reviews
    Price from $9.99 to $1999.99
    transient transfection - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Biophysical properties of AKAP95 protein condensates regulate splicing and tumorigenesis"

    Article Title: Biophysical properties of AKAP95 protein condensates regulate splicing and tumorigenesis

    Journal: bioRxiv

    doi: 10.1101/536839

    AKAP95 condensation requires Tyrosine in 101-210 and regulates splicing. a, Alignment of human and mouse AKAP95 (101-210). Middle row shows identical residues (by letter) and conservative mutations (by “+”). Tyr, red; Phe, blue and tall. Box, Tyr and Phe swapping. b, MBP alone (none) or MBP-AKAP95 (101-210) WT or mutants all at 50 μM and in 30 mM NaCl after TEV protease treatment for 30 min. Turbidity of each reaction was shown in pictures, and by OD600 as mean ± SD from n = 3 (for YA, YS) or 4 (the rest) independent assays. Samples taken after mixing (“mixed”) and from supernatant after centrifugation were resolved by SDS-PAGE followed by coomassie blue staining. c, DIC and fluorescence microscopy images for 10 μM Oregon-green-labeled MBP-AKAP95 (101-210) WT and mutants in 30 mM NaCl after TEV protease treatment for 30 min. Plots from left to right at bottom show the relative protein amount in droplet, number of droplets in a field, and the ratio of protein concentration inside droplets over sum of inside and outside droplets, respectively. Calculated as mean ± SD from n = 24 randomly picked droplets for WT or YF, except for number of droplets from n = 3 randomly picked fields. NA, not applicable. d, Fluorescence microscopy images of HeLa cells transfected with (top) and Flp-In T-Rex 293 cell lines expressing (bottom) GFP fusions with full-length AKAP95 WT or mutants. Repeated > 3 times. e,h, HEK293 cells co-transfected with indicated siRNAs and plasmids were subject to splice reporter assay (top) and immunoblotting with α-AKAP95 (bottom). Δ = Δ(101-210). Mean ± SD from n = 8 [except 5 for Δ(101-210) and 13 for YF and 2 nd WT] independent transfections are plotted in e and 7 independent transfections in h. f, Schematic of AKAP95 chimeras. g, Fluorescence microscopy images of 293T cells transfected with indicated AKAP95 chimeras fused to GFP. Repeated > 3 times. P values by two-sided Student’s t -test for b and one-way ANOVA followed by Tukey’s post hoc test for e and h. Scale bar, 5 μm for all. Uncropped blots are provided as source data.
    Figure Legend Snippet: AKAP95 condensation requires Tyrosine in 101-210 and regulates splicing. a, Alignment of human and mouse AKAP95 (101-210). Middle row shows identical residues (by letter) and conservative mutations (by “+”). Tyr, red; Phe, blue and tall. Box, Tyr and Phe swapping. b, MBP alone (none) or MBP-AKAP95 (101-210) WT or mutants all at 50 μM and in 30 mM NaCl after TEV protease treatment for 30 min. Turbidity of each reaction was shown in pictures, and by OD600 as mean ± SD from n = 3 (for YA, YS) or 4 (the rest) independent assays. Samples taken after mixing (“mixed”) and from supernatant after centrifugation were resolved by SDS-PAGE followed by coomassie blue staining. c, DIC and fluorescence microscopy images for 10 μM Oregon-green-labeled MBP-AKAP95 (101-210) WT and mutants in 30 mM NaCl after TEV protease treatment for 30 min. Plots from left to right at bottom show the relative protein amount in droplet, number of droplets in a field, and the ratio of protein concentration inside droplets over sum of inside and outside droplets, respectively. Calculated as mean ± SD from n = 24 randomly picked droplets for WT or YF, except for number of droplets from n = 3 randomly picked fields. NA, not applicable. d, Fluorescence microscopy images of HeLa cells transfected with (top) and Flp-In T-Rex 293 cell lines expressing (bottom) GFP fusions with full-length AKAP95 WT or mutants. Repeated > 3 times. e,h, HEK293 cells co-transfected with indicated siRNAs and plasmids were subject to splice reporter assay (top) and immunoblotting with α-AKAP95 (bottom). Δ = Δ(101-210). Mean ± SD from n = 8 [except 5 for Δ(101-210) and 13 for YF and 2 nd WT] independent transfections are plotted in e and 7 independent transfections in h. f, Schematic of AKAP95 chimeras. g, Fluorescence microscopy images of 293T cells transfected with indicated AKAP95 chimeras fused to GFP. Repeated > 3 times. P values by two-sided Student’s t -test for b and one-way ANOVA followed by Tukey’s post hoc test for e and h. Scale bar, 5 μm for all. Uncropped blots are provided as source data.

    Techniques Used: Centrifugation, SDS Page, Staining, Fluorescence, Microscopy, Labeling, Protein Concentration, Transfection, Expressing, Reporter Assay

    AKAP95 forms dynamic foci in cell nucleus. a, Immunostaining of endogenous AKAP95 (red) and DNA (DAPI, blue) in indicated cancer cell lines and primary MEFs from WT and Akap95 KO embryos. b, Confocal microscopy images of AKAP95 WT or ZF C-S fused to GFP in nuclei following transfection into HeLa cells. c, Fluorescence microscopy images of HeLa cells transiently expressing AKAP95 WT or Δ(101-210 fused to GFP. d, HeLa cells were transfected with AKAP95-GFP, and two nuclei were imaged at different time points. Time 0 was 24 hr after transfection. Note the growth and merge of the foci, especially those in the red circle. e, Rapid fusion of AKAP95 (ZF C-S )-GFP foci in a HeLa cell nucleus. The white oval and arrow show two different fusion events. These images are from movie 2. All experiments were Repeated > 3 times. Scale bar, 5 μm for all.
    Figure Legend Snippet: AKAP95 forms dynamic foci in cell nucleus. a, Immunostaining of endogenous AKAP95 (red) and DNA (DAPI, blue) in indicated cancer cell lines and primary MEFs from WT and Akap95 KO embryos. b, Confocal microscopy images of AKAP95 WT or ZF C-S fused to GFP in nuclei following transfection into HeLa cells. c, Fluorescence microscopy images of HeLa cells transiently expressing AKAP95 WT or Δ(101-210 fused to GFP. d, HeLa cells were transfected with AKAP95-GFP, and two nuclei were imaged at different time points. Time 0 was 24 hr after transfection. Note the growth and merge of the foci, especially those in the red circle. e, Rapid fusion of AKAP95 (ZF C-S )-GFP foci in a HeLa cell nucleus. The white oval and arrow show two different fusion events. These images are from movie 2. All experiments were Repeated > 3 times. Scale bar, 5 μm for all.

    Techniques Used: Immunostaining, Confocal Microscopy, Transfection, Fluorescence, Microscopy, Expressing

    Related Articles

    Transfection:

    Article Title: Integrative Vectors for Regulated Expression of SARS-CoV-2 Proteins Implicated in RNA Metabolism
    Article Snippet: .. For the transient transfection experiments, cells were induced two hours post-transfection with 1 μg/mL of doxycycline. .. As with the stable cell lines, inductions were staggered and all timepoints were harvested at once.

    Article Title: Constitutive Activity of the Human TRPML2 Channel Induces Cell Degeneration *
    Article Snippet: .. All TRPML constructs for expression in S2 cells were inserted into pMT vectors as described above and transiently expressed using the escort IV transfection reagent (Sigma). .. Human embryonic kidney and HeLa cells were grown in 25-cm2 flasks at 37 °C in Dulbecco's modified Eagle's medium (Biological Industries) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and 1% l -glutamine.

    Article Title: A novel role of vitronectin in promoting survival of mesenchymal stem cells under serum deprivation stress
    Article Snippet: .. To test the efficiency of transient transfection method, 1 μg of a GFP plasmid DNA was used. .. Signal coming from GFP plasmid was visualized using fluorescence microscope.

    Article Title: Evolutionary divergence reveals the molecular basis of EMRE dependence of the human MCU
    Article Snippet: .. 2 d after transient transfection, the cells were DSP cross-linked as described below and lysed as described above. .. A small fraction of the lysate was used to prepare samples at 1 μg/μl of protein concentration in reducing sample buffer for Western blot analysis of lysates.

    Article Title: Improving the flexibility of Genetically Encoded Voltage Indicators via intermolecular FRET
    Article Snippet: .. Transient transfection of cultured mouse hippocampal neurons were done 5-7 days in vitro (DIV) using Lipofectamine 2000 according to the manufacturer’s protocol and experimented on DIV 8–12. .. Equal concentrations of DNA plasmids were used for co-transfection experiments.

    Article Title: miR-125a-5p: a novel regulator of SLC26A6 expression in intestinal epithelial cells
    Article Snippet: .. Transient transfection and 3′-UTR activity. .. Caco-2, HT-29, T-84, and SK-CO15 cells were transiently transfected with 1.5 μg of pmirGLO 3′-UTR of PAT-1 alone or in combination with 20 nM different microRNA mimics/negative control miRNA (Sigma, St. Louis, MO) as previously described us ( ).

    Article Title: Binding of CAP70 to Inducible Nitric Oxide Synthase and Implications for the Vectorial Release of Nitric Oxide in Polarized Cells
    Article Snippet: .. Glutamine, antibiotics, cell culture media (Dulbecco′s modified Eagle's medium), sulfanilic acid, N -(1-naphthyl)-ethylenediamine dihydrochloride, cytochrome-c, oxy-ferrous hemoglobin, nickel-nitrilotriacetic acid resin, transfection reagent Escort-IV, X-β-Gal, cytochalasin-D, oligonucleotides, and Hoechst were purchased from Sigma (St. Louis, MO). .. Trypsin-EDTA and fetal bovine serum were from BioWhittaker Inc. (Walkersville, MD).

    Article Title: Death‐associated protein kinase 1 suppresses hepatocellular carcinoma cell migration and invasion by upregulation of DEAD‐box helicase 20, et al. Death‐associated protein kinase 1 suppresses hepatocellular carcinoma cell migration and invasion by upregulation of DEAD‐box helicase 20
    Article Snippet: .. Transient transfection was carried out using VitalGENE‐II In Vitro DNA Transfection Kit (Biocanaan) according to the manufacturer’s instructions. .. Cells were treated with CHX at 10 μg/mL (Sigma‐Aldrich) for 4 hours, chloroquine at 1 μg/mL (Sigma‐Aldrich) for 24 hours, and MG132 at 1 μg/mL (Sigma‐Aldrich) for 6 hours.

    In Vitro:

    Article Title: Improving the flexibility of Genetically Encoded Voltage Indicators via intermolecular FRET
    Article Snippet: .. Transient transfection of cultured mouse hippocampal neurons were done 5-7 days in vitro (DIV) using Lipofectamine 2000 according to the manufacturer’s protocol and experimented on DIV 8–12. .. Equal concentrations of DNA plasmids were used for co-transfection experiments.

    Article Title: Death‐associated protein kinase 1 suppresses hepatocellular carcinoma cell migration and invasion by upregulation of DEAD‐box helicase 20, et al. Death‐associated protein kinase 1 suppresses hepatocellular carcinoma cell migration and invasion by upregulation of DEAD‐box helicase 20
    Article Snippet: .. Transient transfection was carried out using VitalGENE‐II In Vitro DNA Transfection Kit (Biocanaan) according to the manufacturer’s instructions. .. Cells were treated with CHX at 10 μg/mL (Sigma‐Aldrich) for 4 hours, chloroquine at 1 μg/mL (Sigma‐Aldrich) for 24 hours, and MG132 at 1 μg/mL (Sigma‐Aldrich) for 6 hours.

    Construct:

    Article Title: Constitutive Activity of the Human TRPML2 Channel Induces Cell Degeneration *
    Article Snippet: .. All TRPML constructs for expression in S2 cells were inserted into pMT vectors as described above and transiently expressed using the escort IV transfection reagent (Sigma). .. Human embryonic kidney and HeLa cells were grown in 25-cm2 flasks at 37 °C in Dulbecco's modified Eagle's medium (Biological Industries) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and 1% l -glutamine.

    Activity Assay:

    Article Title: miR-125a-5p: a novel regulator of SLC26A6 expression in intestinal epithelial cells
    Article Snippet: .. Transient transfection and 3′-UTR activity. .. Caco-2, HT-29, T-84, and SK-CO15 cells were transiently transfected with 1.5 μg of pmirGLO 3′-UTR of PAT-1 alone or in combination with 20 nM different microRNA mimics/negative control miRNA (Sigma, St. Louis, MO) as previously described us ( ).

    Cell Culture:

    Article Title: Improving the flexibility of Genetically Encoded Voltage Indicators via intermolecular FRET
    Article Snippet: .. Transient transfection of cultured mouse hippocampal neurons were done 5-7 days in vitro (DIV) using Lipofectamine 2000 according to the manufacturer’s protocol and experimented on DIV 8–12. .. Equal concentrations of DNA plasmids were used for co-transfection experiments.

    Article Title: Binding of CAP70 to Inducible Nitric Oxide Synthase and Implications for the Vectorial Release of Nitric Oxide in Polarized Cells
    Article Snippet: .. Glutamine, antibiotics, cell culture media (Dulbecco′s modified Eagle's medium), sulfanilic acid, N -(1-naphthyl)-ethylenediamine dihydrochloride, cytochrome-c, oxy-ferrous hemoglobin, nickel-nitrilotriacetic acid resin, transfection reagent Escort-IV, X-β-Gal, cytochalasin-D, oligonucleotides, and Hoechst were purchased from Sigma (St. Louis, MO). .. Trypsin-EDTA and fetal bovine serum were from BioWhittaker Inc. (Walkersville, MD).

    Expressing:

    Article Title: Constitutive Activity of the Human TRPML2 Channel Induces Cell Degeneration *
    Article Snippet: .. All TRPML constructs for expression in S2 cells were inserted into pMT vectors as described above and transiently expressed using the escort IV transfection reagent (Sigma). .. Human embryonic kidney and HeLa cells were grown in 25-cm2 flasks at 37 °C in Dulbecco's modified Eagle's medium (Biological Industries) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and 1% l -glutamine.

    Modification:

    Article Title: Binding of CAP70 to Inducible Nitric Oxide Synthase and Implications for the Vectorial Release of Nitric Oxide in Polarized Cells
    Article Snippet: .. Glutamine, antibiotics, cell culture media (Dulbecco′s modified Eagle's medium), sulfanilic acid, N -(1-naphthyl)-ethylenediamine dihydrochloride, cytochrome-c, oxy-ferrous hemoglobin, nickel-nitrilotriacetic acid resin, transfection reagent Escort-IV, X-β-Gal, cytochalasin-D, oligonucleotides, and Hoechst were purchased from Sigma (St. Louis, MO). .. Trypsin-EDTA and fetal bovine serum were from BioWhittaker Inc. (Walkersville, MD).

    Plasmid Preparation:

    Article Title: A novel role of vitronectin in promoting survival of mesenchymal stem cells under serum deprivation stress
    Article Snippet: .. To test the efficiency of transient transfection method, 1 μg of a GFP plasmid DNA was used. .. Signal coming from GFP plasmid was visualized using fluorescence microscope.

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  • 88
    Millipore transfection viral supernatants
    Rescue of anchorage-independent growth in LKPH2 cells with ectopic expression of RNAi-resistant mouse Bub1b cDNAs A. Schematic representation of the mouse Bub1b cDNAs used in this study. Bub1b_FL, full length Bub1b cDNA. Bub1b_∆N, N-terminal truncated Bub1b cDNA. Bub1b_∆K, kinase domain deleted Bub1b cDNA, Bub1b_E406K, full length Bub1b cDNA containing a point mutation (indicated by the red asterisk) in the GLEBS domain that interrupts BUB3 binding and kinetochore localization. The conserved functional motifs are indicated. B and C. BUB1B protein levels in LKPH2 cells stably expressing the indicated cDNAs 72 hours after <t>transfection</t> with siNTC or the indicated siBub1b. Red asterisks indicate the flag-tagged full length (B) or truncated (C) BUB1B. EV: empty vector. D and E. Soft agar colony formation of LKPH2 cells stably expressing the indicated cDNAs transfected with siNTC or the indicated siBub1b. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from three to five independent experiments. **** indicates p
    Transfection Viral Supernatants, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transfection viral supernatants/product/Millipore
    Average 88 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    transfection viral supernatants - by Bioz Stars, 2020-09
    88/100 stars
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    Image Search Results


    Rescue of anchorage-independent growth in LKPH2 cells with ectopic expression of RNAi-resistant mouse Bub1b cDNAs A. Schematic representation of the mouse Bub1b cDNAs used in this study. Bub1b_FL, full length Bub1b cDNA. Bub1b_∆N, N-terminal truncated Bub1b cDNA. Bub1b_∆K, kinase domain deleted Bub1b cDNA, Bub1b_E406K, full length Bub1b cDNA containing a point mutation (indicated by the red asterisk) in the GLEBS domain that interrupts BUB3 binding and kinetochore localization. The conserved functional motifs are indicated. B and C. BUB1B protein levels in LKPH2 cells stably expressing the indicated cDNAs 72 hours after transfection with siNTC or the indicated siBub1b. Red asterisks indicate the flag-tagged full length (B) or truncated (C) BUB1B. EV: empty vector. D and E. Soft agar colony formation of LKPH2 cells stably expressing the indicated cDNAs transfected with siNTC or the indicated siBub1b. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from three to five independent experiments. **** indicates p

    Journal: Genes & Cancer

    Article Title: Requirement for BUB1B/BUBR1 in tumor progression of lung adenocarcinoma

    doi:

    Figure Lengend Snippet: Rescue of anchorage-independent growth in LKPH2 cells with ectopic expression of RNAi-resistant mouse Bub1b cDNAs A. Schematic representation of the mouse Bub1b cDNAs used in this study. Bub1b_FL, full length Bub1b cDNA. Bub1b_∆N, N-terminal truncated Bub1b cDNA. Bub1b_∆K, kinase domain deleted Bub1b cDNA, Bub1b_E406K, full length Bub1b cDNA containing a point mutation (indicated by the red asterisk) in the GLEBS domain that interrupts BUB3 binding and kinetochore localization. The conserved functional motifs are indicated. B and C. BUB1B protein levels in LKPH2 cells stably expressing the indicated cDNAs 72 hours after transfection with siNTC or the indicated siBub1b. Red asterisks indicate the flag-tagged full length (B) or truncated (C) BUB1B. EV: empty vector. D and E. Soft agar colony formation of LKPH2 cells stably expressing the indicated cDNAs transfected with siNTC or the indicated siBub1b. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from three to five independent experiments. **** indicates p

    Article Snippet: Forty-eight hours after transfection viral supernatants were collected and used to infect LKPH2 cells in the presence of 8μg/mL polybrene (EMD Millipore Corp).

    Techniques: Expressing, Mutagenesis, Binding Assay, Functional Assay, Stable Transfection, Transfection, Plasmid Preparation