lipid mediated transfection protocol  (Thermo Fisher)


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    Name:
    Lipofectamine 2000 Transfection Reagent
    Description:
    Lipofectamine 2000 Transfection Reagent is a versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines Researchers use Lipofectamine 2000 Reagent for siRNA and shRNA based gene knockdown experiments as well as for gene expression studies With Lipofectamine 2000 Transfection Reagent you ll get • Exceptional transfection efficiency in the broadest range of cell lines and the highest levels of recombinant protein expression see table • Superior performance for co transfection of siRNA and plasmid DNA• Proven efficacy in the presence of serum eliminates the need to change media following transfection• Reliable performance for high throughput applications• The best choice for establishing stable cell linesA high performance transfection reagent for gene expression and gene silencingLipofectamine 2000 Transfection Reagent works effectively with all common cell lines as well as with many challenging ones and can be used in media with or without serum For gene silencing Lipofectamine 2000 Transfection Reagent s high efficiency transfections provide the high levels of gene knockdown needed to produce convincing results Lipofectamine 2000 Transfection Reagent is the number one choice for co transfection given its effectiveness for transfecting both siRNA and plasmid DNA Lipofectamine 2000 Transfection Reagent is easy to use simply mix with nucleic acid and add to cell culture Ideal for high throughput workSimplicity and speed combined with high transfection efficiency make Lipofectamine 2000 Transfection Reagent ideal for transient protein expression or high throughput RNAi transfections Transfection conditions can be easily established for automated or robotic systems used in such applications
    Catalog Number:
    11668019
    Price:
    None
    Applications:
    Cell Culture|Plasmid Transfection|RNAi Transfection|Stem Cell & Primary Cell Transfections|Synthetic siRNA Transfection|shRNA & miR RNAi Plasmid Transfection|Transfection
    Category:
    Cell Culture Transfection Reagents
    Buy from Supplier


    Structured Review

    Thermo Fisher lipid mediated transfection protocol
    Differential expression is abrogated with <t>transfection,</t> time in culture, and age of DNA. Represents the fold difference in transcriptional activity after normalizing luciferase RLUs/ to −308G/GL3Luc, which was set at 1. ( a ) Jurkat cells were transiently
    Lipofectamine 2000 Transfection Reagent is a versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines Researchers use Lipofectamine 2000 Reagent for siRNA and shRNA based gene knockdown experiments as well as for gene expression studies With Lipofectamine 2000 Transfection Reagent you ll get • Exceptional transfection efficiency in the broadest range of cell lines and the highest levels of recombinant protein expression see table • Superior performance for co transfection of siRNA and plasmid DNA• Proven efficacy in the presence of serum eliminates the need to change media following transfection• Reliable performance for high throughput applications• The best choice for establishing stable cell linesA high performance transfection reagent for gene expression and gene silencingLipofectamine 2000 Transfection Reagent works effectively with all common cell lines as well as with many challenging ones and can be used in media with or without serum For gene silencing Lipofectamine 2000 Transfection Reagent s high efficiency transfections provide the high levels of gene knockdown needed to produce convincing results Lipofectamine 2000 Transfection Reagent is the number one choice for co transfection given its effectiveness for transfecting both siRNA and plasmid DNA Lipofectamine 2000 Transfection Reagent is easy to use simply mix with nucleic acid and add to cell culture Ideal for high throughput workSimplicity and speed combined with high transfection efficiency make Lipofectamine 2000 Transfection Reagent ideal for transient protein expression or high throughput RNAi transfections Transfection conditions can be easily established for automated or robotic systems used in such applications
    https://www.bioz.com/result/lipid mediated transfection protocol/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    lipid mediated transfection protocol - by Bioz Stars, 2020-09
    99/100 stars

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    Images

    1) Product Images from "A critical assessment of the factors affecting reporter gene assays for promoter SNP function: a reassessment of -308 TNF polymorphism function using a novel integrated reporter system"

    Article Title: A critical assessment of the factors affecting reporter gene assays for promoter SNP function: a reassessment of -308 TNF polymorphism function using a novel integrated reporter system

    Journal: European Journal of Human Genetics

    doi: 10.1038/ejhg.2009.80

    Differential expression is abrogated with transfection, time in culture, and age of DNA. Represents the fold difference in transcriptional activity after normalizing luciferase RLUs/ to −308G/GL3Luc, which was set at 1. ( a ) Jurkat cells were transiently
    Figure Legend Snippet: Differential expression is abrogated with transfection, time in culture, and age of DNA. Represents the fold difference in transcriptional activity after normalizing luciferase RLUs/ to −308G/GL3Luc, which was set at 1. ( a ) Jurkat cells were transiently

    Techniques Used: Expressing, Transfection, Activity Assay, Luciferase

    Effects of co-transfection with pRL-TK control plasmid. Represents the fold difference in transcriptional activity of −308A/GL3Luc and −308G/GL3Luc after normalizing RLU/s to −308G/GL3Luc only, which was set at 1. Jurkat cells
    Figure Legend Snippet: Effects of co-transfection with pRL-TK control plasmid. Represents the fold difference in transcriptional activity of −308A/GL3Luc and −308G/GL3Luc after normalizing RLU/s to −308G/GL3Luc only, which was set at 1. Jurkat cells

    Techniques Used: Cotransfection, Plasmid Preparation, Activity Assay

    Related Articles

    MTT Assay:

    Article Title: Influence of Oligospermines Architecture on Their Suitability for siRNA Delivery
    Article Snippet: .. Lipofectamine 2000 (LF), SYBR Gold dye, and (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Life Technologies (Grand Island, NY). .. Polyethylenimine (PEI, MW 5 kDa) was obtained from BASF (Lupasol, Cologne, Germany).

    Transfection:

    Article Title: Knockdown of Dinoflagellate Cellulose Synthase CesA1 Resulted in Malformed Intracellular Cellulosic Thecal Plates and Severely Impeded Cyst-to-Swarmer Transition
    Article Snippet: .. For each transfection sample, oligomer-Lipofectamine 2000 complexes were prepared by mixing 300 μl L medium, 300 μl 12 nmole antisense or scrambled ODN and 30 μl Lipofectamine Reagent 2000 (Invitrogen). .. The mixture was incubated for 30 min at room temperature (with rotation), before incubating with 300 μl spheroplasts (original culture: 800 ml, final cell number: ∼1.8 × 106 ) for another 1 h at room temperature (with gentle rotation).

    Article Title: Estrogen Receptor β Agonists Differentially Affect the Growth of Human Melanoma Cell Lines
    Article Snippet: .. ERβ transcriptional activity assay BLM cells, seeded in 24-well plates (5x104 cells/well) in phenol red free DMEM medium supplemented with 10% charcoal stripped FBS, were transfected using Lipofectamine 2000 (Life Technologies, Monza, Italy), according to the manufacturer’s protocol [ ]. .. The following constructs were cotransfected: pVERE-tk-Luc (1μg), the reporter plasmid encoding the firefly luciferase reporter gene under the control of the estrogen response element (ERE; kindly provided by Dr. A. Maggi, Department of Pharmacological and Biomolecular Sciences, University of Milano, Milano, Italy), to evaluate the transcriptional activity of ERβ, and pCMVβ (0.4 μg), the reporter plasmid encoding the β-galactosidase (Clontech, Jesi, Italy), as the internal control plasmid.

    Article Title: Nanoparticle-Mediated Gene Silencing for Sensitization of Lung Cancer to Cisplatin Therapy
    Article Snippet: .. Negative controls consisted of untreated cells or cells transfected with control scrambled siRNA while positive control cells were transfected with Lipofectamine 2000 (Life Technologies) lipoplexes, which were prepared as described above. .. Unless otherwise stated, cells were transfected twice at 24 h intervals in 37 °C and 5% CO2 with 200 μL of micelleplex suspension containing 100 pmol siRNA within a total volume of 2 mL of serum-containing cell culture media.

    Article Title: Mitochondrial-targeted Aryl Hydrocarbon Receptor and the Impact of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin on Cellular Respiration and the Mitochondrial Proteome
    Article Snippet: .. When cells were 50% confluent, the siRNAs specific for AIP (Ambion s62179 (siAIP1), s62181 (siAIP2), Life Technologies), and the Silencer® Select Negative Control no. 1 siRNA (Ambion) were transfected into hepa1c1c7 cells using Lipofectamine 2000 (Invitrogen) following the manufacturer's protocol. .. For TOMM20 knockdown, the siRNA for TOMM20 (Tomm20 ON-TARGET plus, SMARTpool L-006487-01-0005, Dharmacon, GE) and the nontargeting siRNA (Dharmacon, GE) were transfected into hepa1c1c7 cells.

    Recombinant:

    Article Title: Chemotherapy-induced pyroptosis is mediated by BAK/BAX-caspase-3-GSDME pathway and inhibited by 2-bromopalmitate
    Article Snippet: .. Reagents were purchased as followings: recombinant human TNFα (rcyc-htnfs) from InvivoGene; cycloheximide (HY-12320), navitoclax (HY-10087), actinomycin D(HY-17559), broad spectrum caspase inhibitor Q-VD-OPh (HY-12305) from MedChemExpress; caspase-3 inhitor Q-DEVD-OPh (1175-1) from Biovision, protease inhibitor cocktail tablets (04693132001) from Roche; protein G sepharose (17-0618-01) from GE Healthcare; Lipofectamine 2000 (11668-019) and Lipofectamine RNAiMAX (13778-150) from Invitrogen; anti-Flag M2 magnetic beads (M8823) and 2-BP (238422) from Sigma-Aldrich; CytoTox 96 Non-Radio cytotoxicity assay kit (G1780) from Promega; Hoechst 33342/PI double stain kit (CA1120) from Solarbio; and FITC annexin V appotosis kit I (556547) from BD Biosciences. .. Cell culture and treatments The human colon cancer cell lines HCT116 wild type and BAK −/− BAX −/− (DKO), the mouse embryonic fibroblasts MEFs, the human ovarian cancer cell lines including Ovcar3, Ovcar5, Ovcar8, A2780, HeyA8, OV90, COV362, SKOV3, Ovise, and the hematoma cell lines including V937, MOLT-3, MOLT-4, THP-1, SKW6.4, H9, Jurkat, K562, KG1a, RL, ML-1, HL-60, HT, and MV4-11 were kind gifts from Scott H. Kaufmann (Mayo Clinic, Rochester, MN).

    Positive Control:

    Article Title: Nanoparticle-Mediated Gene Silencing for Sensitization of Lung Cancer to Cisplatin Therapy
    Article Snippet: .. Negative controls consisted of untreated cells or cells transfected with control scrambled siRNA while positive control cells were transfected with Lipofectamine 2000 (Life Technologies) lipoplexes, which were prepared as described above. .. Unless otherwise stated, cells were transfected twice at 24 h intervals in 37 °C and 5% CO2 with 200 μL of micelleplex suspension containing 100 pmol siRNA within a total volume of 2 mL of serum-containing cell culture media.

    Magnetic Beads:

    Article Title: Chemotherapy-induced pyroptosis is mediated by BAK/BAX-caspase-3-GSDME pathway and inhibited by 2-bromopalmitate
    Article Snippet: .. Reagents were purchased as followings: recombinant human TNFα (rcyc-htnfs) from InvivoGene; cycloheximide (HY-12320), navitoclax (HY-10087), actinomycin D(HY-17559), broad spectrum caspase inhibitor Q-VD-OPh (HY-12305) from MedChemExpress; caspase-3 inhitor Q-DEVD-OPh (1175-1) from Biovision, protease inhibitor cocktail tablets (04693132001) from Roche; protein G sepharose (17-0618-01) from GE Healthcare; Lipofectamine 2000 (11668-019) and Lipofectamine RNAiMAX (13778-150) from Invitrogen; anti-Flag M2 magnetic beads (M8823) and 2-BP (238422) from Sigma-Aldrich; CytoTox 96 Non-Radio cytotoxicity assay kit (G1780) from Promega; Hoechst 33342/PI double stain kit (CA1120) from Solarbio; and FITC annexin V appotosis kit I (556547) from BD Biosciences. .. Cell culture and treatments The human colon cancer cell lines HCT116 wild type and BAK −/− BAX −/− (DKO), the mouse embryonic fibroblasts MEFs, the human ovarian cancer cell lines including Ovcar3, Ovcar5, Ovcar8, A2780, HeyA8, OV90, COV362, SKOV3, Ovise, and the hematoma cell lines including V937, MOLT-3, MOLT-4, THP-1, SKW6.4, H9, Jurkat, K562, KG1a, RL, ML-1, HL-60, HT, and MV4-11 were kind gifts from Scott H. Kaufmann (Mayo Clinic, Rochester, MN).

    Protease Inhibitor:

    Article Title: Chemotherapy-induced pyroptosis is mediated by BAK/BAX-caspase-3-GSDME pathway and inhibited by 2-bromopalmitate
    Article Snippet: .. Reagents were purchased as followings: recombinant human TNFα (rcyc-htnfs) from InvivoGene; cycloheximide (HY-12320), navitoclax (HY-10087), actinomycin D(HY-17559), broad spectrum caspase inhibitor Q-VD-OPh (HY-12305) from MedChemExpress; caspase-3 inhitor Q-DEVD-OPh (1175-1) from Biovision, protease inhibitor cocktail tablets (04693132001) from Roche; protein G sepharose (17-0618-01) from GE Healthcare; Lipofectamine 2000 (11668-019) and Lipofectamine RNAiMAX (13778-150) from Invitrogen; anti-Flag M2 magnetic beads (M8823) and 2-BP (238422) from Sigma-Aldrich; CytoTox 96 Non-Radio cytotoxicity assay kit (G1780) from Promega; Hoechst 33342/PI double stain kit (CA1120) from Solarbio; and FITC annexin V appotosis kit I (556547) from BD Biosciences. .. Cell culture and treatments The human colon cancer cell lines HCT116 wild type and BAK −/− BAX −/− (DKO), the mouse embryonic fibroblasts MEFs, the human ovarian cancer cell lines including Ovcar3, Ovcar5, Ovcar8, A2780, HeyA8, OV90, COV362, SKOV3, Ovise, and the hematoma cell lines including V937, MOLT-3, MOLT-4, THP-1, SKW6.4, H9, Jurkat, K562, KG1a, RL, ML-1, HL-60, HT, and MV4-11 were kind gifts from Scott H. Kaufmann (Mayo Clinic, Rochester, MN).

    Negative Control:

    Article Title: Mitochondrial-targeted Aryl Hydrocarbon Receptor and the Impact of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin on Cellular Respiration and the Mitochondrial Proteome
    Article Snippet: .. When cells were 50% confluent, the siRNAs specific for AIP (Ambion s62179 (siAIP1), s62181 (siAIP2), Life Technologies), and the Silencer® Select Negative Control no. 1 siRNA (Ambion) were transfected into hepa1c1c7 cells using Lipofectamine 2000 (Invitrogen) following the manufacturer's protocol. .. For TOMM20 knockdown, the siRNA for TOMM20 (Tomm20 ON-TARGET plus, SMARTpool L-006487-01-0005, Dharmacon, GE) and the nontargeting siRNA (Dharmacon, GE) were transfected into hepa1c1c7 cells.

    Mouse Assay:

    Article Title: Cholesterol-Containing Nuclease-Resistant siRNA Accumulates in Tumors in a Carrier-free Mode and Silences MDR1 Gene
    Article Snippet: .. In each experiment, four tumor-bearing or healthy mice were injected intravenously (i.v.), intraperitoneally (i.p.), intramuscularly (i.m.), subcutaneously (s.c.), or peritumorally (p.t.) with 1.7 μg/g Cy7 or Cy5.5-labeled siRNA, Ch-siRNA, or siRNA/Lipofectamine 2000 complex in 100 μL Opti-MEM medium (Invitrogen), and the fifth mouse was left intact as a control. ..

    Incubation:

    Article Title: Aβ and tau prion-like activities decline with longevity in Alzheimer’s disease brains
    Article Snippet: .. A mixture of Lipofectamine 2000 (1.5% final volume; Thermo Fisher), OptiMEM (78.5% final volume; Thermo Fisher) and sample (20% final volume) were incubated at room temperature for 2 hours and plated in four replicate wells (10 μL/well). .. Plates were then incubated and imaged every 24 hours on the IN Cell 6000 (GE Healthcare) for 3–4 days.

    Activity Assay:

    Article Title: Estrogen Receptor β Agonists Differentially Affect the Growth of Human Melanoma Cell Lines
    Article Snippet: .. ERβ transcriptional activity assay BLM cells, seeded in 24-well plates (5x104 cells/well) in phenol red free DMEM medium supplemented with 10% charcoal stripped FBS, were transfected using Lipofectamine 2000 (Life Technologies, Monza, Italy), according to the manufacturer’s protocol [ ]. .. The following constructs were cotransfected: pVERE-tk-Luc (1μg), the reporter plasmid encoding the firefly luciferase reporter gene under the control of the estrogen response element (ERE; kindly provided by Dr. A. Maggi, Department of Pharmacological and Biomolecular Sciences, University of Milano, Milano, Italy), to evaluate the transcriptional activity of ERβ, and pCMVβ (0.4 μg), the reporter plasmid encoding the β-galactosidase (Clontech, Jesi, Italy), as the internal control plasmid.

    Staining:

    Article Title: Chemotherapy-induced pyroptosis is mediated by BAK/BAX-caspase-3-GSDME pathway and inhibited by 2-bromopalmitate
    Article Snippet: .. Reagents were purchased as followings: recombinant human TNFα (rcyc-htnfs) from InvivoGene; cycloheximide (HY-12320), navitoclax (HY-10087), actinomycin D(HY-17559), broad spectrum caspase inhibitor Q-VD-OPh (HY-12305) from MedChemExpress; caspase-3 inhitor Q-DEVD-OPh (1175-1) from Biovision, protease inhibitor cocktail tablets (04693132001) from Roche; protein G sepharose (17-0618-01) from GE Healthcare; Lipofectamine 2000 (11668-019) and Lipofectamine RNAiMAX (13778-150) from Invitrogen; anti-Flag M2 magnetic beads (M8823) and 2-BP (238422) from Sigma-Aldrich; CytoTox 96 Non-Radio cytotoxicity assay kit (G1780) from Promega; Hoechst 33342/PI double stain kit (CA1120) from Solarbio; and FITC annexin V appotosis kit I (556547) from BD Biosciences. .. Cell culture and treatments The human colon cancer cell lines HCT116 wild type and BAK −/− BAX −/− (DKO), the mouse embryonic fibroblasts MEFs, the human ovarian cancer cell lines including Ovcar3, Ovcar5, Ovcar8, A2780, HeyA8, OV90, COV362, SKOV3, Ovise, and the hematoma cell lines including V937, MOLT-3, MOLT-4, THP-1, SKW6.4, H9, Jurkat, K562, KG1a, RL, ML-1, HL-60, HT, and MV4-11 were kind gifts from Scott H. Kaufmann (Mayo Clinic, Rochester, MN).

    Injection:

    Article Title: Cholesterol-Containing Nuclease-Resistant siRNA Accumulates in Tumors in a Carrier-free Mode and Silences MDR1 Gene
    Article Snippet: .. In each experiment, four tumor-bearing or healthy mice were injected intravenously (i.v.), intraperitoneally (i.p.), intramuscularly (i.m.), subcutaneously (s.c.), or peritumorally (p.t.) with 1.7 μg/g Cy7 or Cy5.5-labeled siRNA, Ch-siRNA, or siRNA/Lipofectamine 2000 complex in 100 μL Opti-MEM medium (Invitrogen), and the fifth mouse was left intact as a control. ..

    Cytotoxicity Assay:

    Article Title: Chemotherapy-induced pyroptosis is mediated by BAK/BAX-caspase-3-GSDME pathway and inhibited by 2-bromopalmitate
    Article Snippet: .. Reagents were purchased as followings: recombinant human TNFα (rcyc-htnfs) from InvivoGene; cycloheximide (HY-12320), navitoclax (HY-10087), actinomycin D(HY-17559), broad spectrum caspase inhibitor Q-VD-OPh (HY-12305) from MedChemExpress; caspase-3 inhitor Q-DEVD-OPh (1175-1) from Biovision, protease inhibitor cocktail tablets (04693132001) from Roche; protein G sepharose (17-0618-01) from GE Healthcare; Lipofectamine 2000 (11668-019) and Lipofectamine RNAiMAX (13778-150) from Invitrogen; anti-Flag M2 magnetic beads (M8823) and 2-BP (238422) from Sigma-Aldrich; CytoTox 96 Non-Radio cytotoxicity assay kit (G1780) from Promega; Hoechst 33342/PI double stain kit (CA1120) from Solarbio; and FITC annexin V appotosis kit I (556547) from BD Biosciences. .. Cell culture and treatments The human colon cancer cell lines HCT116 wild type and BAK −/− BAX −/− (DKO), the mouse embryonic fibroblasts MEFs, the human ovarian cancer cell lines including Ovcar3, Ovcar5, Ovcar8, A2780, HeyA8, OV90, COV362, SKOV3, Ovise, and the hematoma cell lines including V937, MOLT-3, MOLT-4, THP-1, SKW6.4, H9, Jurkat, K562, KG1a, RL, ML-1, HL-60, HT, and MV4-11 were kind gifts from Scott H. Kaufmann (Mayo Clinic, Rochester, MN).

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  • 88
    Thermo Fisher lipid based transfection reagents
    Lipid Based Transfection Reagents, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipid based transfection reagents/product/Thermo Fisher
    Average 88 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    lipid based transfection reagents - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

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