lipofectamine 2000 cd transfection reagent  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Lipofectamine 2000 CD Transfection Reagent
    Description:
    Lipofectamine 2000 CD Chemically Defined Transfection Reagent is a proprietary animal origin free formulation for transfecting nucleic acids into eukaryotic cells Using Lipofectamine 2000 CD reagent for transfection provides the following advantages • Highest transfection efficiency in many cell types and formats • The animal origin free formulation ensures that mammalian cell culture and bioproduction processes are free of animal derived materials • DNA Lipofectamine 2000 CD complexes can be added directly to cells in culture medium • It is not necessary to remove complexes or change add medium after transfection but complexes may be removed after 4 6 hours
    Catalog Number:
    12566014
    Price:
    None
    Applications:
    Bioproduction|Cell Culture|Plasmid Transfection|Transient Protein Production & High-Throughput Screening|Antibody & Cell Culture Production|Transfection
    Category:
    Cell Culture Transfection Reagents
    Buy from Supplier


    Structured Review

    Thermo Fisher lipofectamine 2000 cd transfection reagent
    Human PBMCs stimulated with DAMPs induce miRNA-30e and enhance innate immune responses: PBMCs from three healthy individuals were transfected with their own genomic DNA (ds DNA). (A) Isolated genomic DNA sonicated into small fragments of dsDNA of approximately 100-150 bps each (as shown) and transfected using Lipofectamine 2000. (B-F) Quantification (by qRT-PCR analysis) of the fold changes in the relative abundances of (B) miR-30e, (C-E) respective innate immune transcripts ( IFNα, IFNβ, IFIT1  and  IL6)  in all individuals and (F) NDV viral transcript (shown as per schematic workflow). Data are mean +/-SEM of triplicate samples from single experiment and are representative of three independent experiments in three different individuals. *** P
    Lipofectamine 2000 CD Chemically Defined Transfection Reagent is a proprietary animal origin free formulation for transfecting nucleic acids into eukaryotic cells Using Lipofectamine 2000 CD reagent for transfection provides the following advantages • Highest transfection efficiency in many cell types and formats • The animal origin free formulation ensures that mammalian cell culture and bioproduction processes are free of animal derived materials • DNA Lipofectamine 2000 CD complexes can be added directly to cells in culture medium • It is not necessary to remove complexes or change add medium after transfection but complexes may be removed after 4 6 hours
    https://www.bioz.com/result/lipofectamine 2000 cd transfection reagent/product/Thermo Fisher
    Average 99 stars, based on 49 article reviews
    Price from $9.99 to $1999.99
    lipofectamine 2000 cd transfection reagent - by Bioz Stars, 2020-11
    99/100 stars

    Images

    1) Product Images from "Integrated role of microRNA-30e-5p through targeting negative regulators of innate immune pathways during HBV infection and SLE"

    Article Title: Integrated role of microRNA-30e-5p through targeting negative regulators of innate immune pathways during HBV infection and SLE

    Journal: bioRxiv

    doi: 10.1101/2020.02.29.969014

    Human PBMCs stimulated with DAMPs induce miRNA-30e and enhance innate immune responses: PBMCs from three healthy individuals were transfected with their own genomic DNA (ds DNA). (A) Isolated genomic DNA sonicated into small fragments of dsDNA of approximately 100-150 bps each (as shown) and transfected using Lipofectamine 2000. (B-F) Quantification (by qRT-PCR analysis) of the fold changes in the relative abundances of (B) miR-30e, (C-E) respective innate immune transcripts ( IFNα, IFNβ, IFIT1  and  IL6)  in all individuals and (F) NDV viral transcript (shown as per schematic workflow). Data are mean +/-SEM of triplicate samples from single experiment and are representative of three independent experiments in three different individuals. *** P
    Figure Legend Snippet: Human PBMCs stimulated with DAMPs induce miRNA-30e and enhance innate immune responses: PBMCs from three healthy individuals were transfected with their own genomic DNA (ds DNA). (A) Isolated genomic DNA sonicated into small fragments of dsDNA of approximately 100-150 bps each (as shown) and transfected using Lipofectamine 2000. (B-F) Quantification (by qRT-PCR analysis) of the fold changes in the relative abundances of (B) miR-30e, (C-E) respective innate immune transcripts ( IFNα, IFNβ, IFIT1 and IL6) in all individuals and (F) NDV viral transcript (shown as per schematic workflow). Data are mean +/-SEM of triplicate samples from single experiment and are representative of three independent experiments in three different individuals. *** P

    Techniques Used: Transfection, Isolation, Sonication, Quantitative RT-PCR

    2) Product Images from "Mitochondrial DNA Stress Signalling Protects the Nuclear Genome"

    Article Title: Mitochondrial DNA Stress Signalling Protects the Nuclear Genome

    Journal: Nature metabolism

    doi: 10.1038/s42255-019-0150-8

    Innate immune signalling by chronic mtDNA stress requires U-ISGF3 but is not associated with NF-κB or interferon gene activation. a-c, qRT-PCR analysis of ( a ) the indicated ISGs, ( b ) NF-κB target genes and (c) IFN genes in WT and Tfam +/− littermate MEFs. d , Western blot of STAT1, p-STAT1 (Y701), TFAM and GAPDH (loading control) in WT and Tfam +/− littermate MEFs transfected with 2μg Poly I:C or lipofectamine only (Mock) for 12 hours. (n=3 independent experiments) e, qRT-PCR analysis of the indicated ISGs in WT, Tfam +/− , Stat1 −/− and Tfam +/− Stat1 −/− littermate MEFs. f, Western blot of STAT1, p-STAT1 (Y701) and histone H3 (nuclear marker and loading control) in purified nuclear extracts from WT and Tfam +/− littermate MEFs. (n=3 independent experiments). g, qRT-PCR analysis of the indicated ISGs in Tfam +/− MEFs transfected with siRNA against Irf9 or Stat2 . All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: **p
    Figure Legend Snippet: Innate immune signalling by chronic mtDNA stress requires U-ISGF3 but is not associated with NF-κB or interferon gene activation. a-c, qRT-PCR analysis of ( a ) the indicated ISGs, ( b ) NF-κB target genes and (c) IFN genes in WT and Tfam +/− littermate MEFs. d , Western blot of STAT1, p-STAT1 (Y701), TFAM and GAPDH (loading control) in WT and Tfam +/− littermate MEFs transfected with 2μg Poly I:C or lipofectamine only (Mock) for 12 hours. (n=3 independent experiments) e, qRT-PCR analysis of the indicated ISGs in WT, Tfam +/− , Stat1 −/− and Tfam +/− Stat1 −/− littermate MEFs. f, Western blot of STAT1, p-STAT1 (Y701) and histone H3 (nuclear marker and loading control) in purified nuclear extracts from WT and Tfam +/− littermate MEFs. (n=3 independent experiments). g, qRT-PCR analysis of the indicated ISGs in Tfam +/− MEFs transfected with siRNA against Irf9 or Stat2 . All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: **p

    Techniques Used: Activation Assay, Quantitative RT-PCR, Western Blot, Transfection, Marker, Purification, Two Tailed Test

    Additional analysis of the STAT1 null (Stat1 −/− ) condition in WT (Tfam +/+ Stat1 +/+ ) and Tfam +/− MEFs. a, Western blot of STAT1, TFAM and GAPDH (loading control) in littermate MEFs of the indicated Tfam and Stat1 genotypes (bottom) that were transfected with 2 μg of Poly(I:C) or lipofectamine only (Mock) for 12 hours. (n=3 independent experiments). MEFs described in a were analysed (all normalized to WT) for b, mtDNA abundance (relative mtDNA copy number) by qPCR with D-loop primers; c, mitochondrial mass using MitoTracker Green (MTG) and flow cytometry (mean fluorescence intensity, MFI, in arbitrary units, A.U., is plotted), and d, mitochondrial membrane potential using MitoTracker Deep Red (MTDR) and flow cytometry (MFI in A.U. is plotted). e, MEFs of the indicated genotypes were analysed by immunofluorescence for mitochondrial and mtDNA nucleoid morphology using antibodies against HSP60 (Mito., magenta) and DNA (DNA, cyan), respectively (n=3 independent experiments). Images are Z-stack projections and scale bar represents 10 μm. f, qRT-PCR analysis of the ISGs Cxcl10 and Ifit1 in MEFs of the indicated genotypes. g, qRT-PCR analysis of the indicated ISGs in Tfam +/− MEFs transfected with control (Ctrl) or Irf3 siRNAs for 72 hours. h, qRT-PCR analysis of the indicated ISGs in Tfam +/− Stat1 −/− MEFs transfected with control (Ctrl) or Irf3 siRNAs for 72 hours. i, Western blot showing siRNA knock-down of IRF9 or STAT2 in Tfam +/− MEFs compared to control (Ctrl) siRNA treated cells. GAPDH was probed as the loading control. (n=3 independent experiments). j, qRT-PCR analysis of the indicated ISGs in Tfam +/− Stat1 −/− MEFs transfected with control (Ctrl) or Irf9 siRNAs for 72 hours. k, qRT-PCR analysis of the indicated ISGs in Tfam +/− Stat1 −/− MEFs transfected with control (Ctrl) or Stat2 siRNAs for 72 hours. c-d, error bars indicate mean ± SD of n=3 biological replicates. For b the data shown are from one of two biological replicates with the error bars indicating the mean ± SD of three technical replicates. For g, h, j and k, . Asterisks indicate significance as follows: ** P
    Figure Legend Snippet: Additional analysis of the STAT1 null (Stat1 −/− ) condition in WT (Tfam +/+ Stat1 +/+ ) and Tfam +/− MEFs. a, Western blot of STAT1, TFAM and GAPDH (loading control) in littermate MEFs of the indicated Tfam and Stat1 genotypes (bottom) that were transfected with 2 μg of Poly(I:C) or lipofectamine only (Mock) for 12 hours. (n=3 independent experiments). MEFs described in a were analysed (all normalized to WT) for b, mtDNA abundance (relative mtDNA copy number) by qPCR with D-loop primers; c, mitochondrial mass using MitoTracker Green (MTG) and flow cytometry (mean fluorescence intensity, MFI, in arbitrary units, A.U., is plotted), and d, mitochondrial membrane potential using MitoTracker Deep Red (MTDR) and flow cytometry (MFI in A.U. is plotted). e, MEFs of the indicated genotypes were analysed by immunofluorescence for mitochondrial and mtDNA nucleoid morphology using antibodies against HSP60 (Mito., magenta) and DNA (DNA, cyan), respectively (n=3 independent experiments). Images are Z-stack projections and scale bar represents 10 μm. f, qRT-PCR analysis of the ISGs Cxcl10 and Ifit1 in MEFs of the indicated genotypes. g, qRT-PCR analysis of the indicated ISGs in Tfam +/− MEFs transfected with control (Ctrl) or Irf3 siRNAs for 72 hours. h, qRT-PCR analysis of the indicated ISGs in Tfam +/− Stat1 −/− MEFs transfected with control (Ctrl) or Irf3 siRNAs for 72 hours. i, Western blot showing siRNA knock-down of IRF9 or STAT2 in Tfam +/− MEFs compared to control (Ctrl) siRNA treated cells. GAPDH was probed as the loading control. (n=3 independent experiments). j, qRT-PCR analysis of the indicated ISGs in Tfam +/− Stat1 −/− MEFs transfected with control (Ctrl) or Irf9 siRNAs for 72 hours. k, qRT-PCR analysis of the indicated ISGs in Tfam +/− Stat1 −/− MEFs transfected with control (Ctrl) or Stat2 siRNAs for 72 hours. c-d, error bars indicate mean ± SD of n=3 biological replicates. For b the data shown are from one of two biological replicates with the error bars indicating the mean ± SD of three technical replicates. For g, h, j and k, . Asterisks indicate significance as follows: ** P

    Techniques Used: Western Blot, Transfection, Real-time Polymerase Chain Reaction, Flow Cytometry, Fluorescence, Immunofluorescence, Quantitative RT-PCR

    Additional analysis of innate immune signalling in MEFs. a, Heat map of normalized expression values of the indicated ISGs (red font), interferon genes (black font), and NF-κB target genes (blue font) from our previously published microarray analysis4 of WT and Tfam+/− MEFs (two of each). b-d, qRT-PCR analyses of the indicated ISGs (b) , NF-κB target genes (c) and interferon genes (d) in WT MEFs transfected with control (Ctrl) or Tfam siRNA for 72 hours. e, Western blot probing the indicated NF-κB pathway proteins, TFAM and VDAC (loading control) in WT and Tfam+/− littermate MEFs. (n=3 independent experiments.) f, qRT-PCR analysis of interferon β (Ifnb) gene expression in WT MEFs transfected with 2 μg poly(I:C) or lipofectamine only (Mock) for 9 hours. g, qRT-PCR analysis of the indicated ISGs in WT MEFs cultured overnight in the presence of control media (plain), media conditioned by WT or Tfam+/− MEFs, or media conditioned by WT MEFs stimulated with poly(I:C) for 9 hours (PIC-stimulated). The data shown are from one of two ( f and g ) or three ( b-d . All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: ** P
    Figure Legend Snippet: Additional analysis of innate immune signalling in MEFs. a, Heat map of normalized expression values of the indicated ISGs (red font), interferon genes (black font), and NF-κB target genes (blue font) from our previously published microarray analysis4 of WT and Tfam+/− MEFs (two of each). b-d, qRT-PCR analyses of the indicated ISGs (b) , NF-κB target genes (c) and interferon genes (d) in WT MEFs transfected with control (Ctrl) or Tfam siRNA for 72 hours. e, Western blot probing the indicated NF-κB pathway proteins, TFAM and VDAC (loading control) in WT and Tfam+/− littermate MEFs. (n=3 independent experiments.) f, qRT-PCR analysis of interferon β (Ifnb) gene expression in WT MEFs transfected with 2 μg poly(I:C) or lipofectamine only (Mock) for 9 hours. g, qRT-PCR analysis of the indicated ISGs in WT MEFs cultured overnight in the presence of control media (plain), media conditioned by WT or Tfam+/− MEFs, or media conditioned by WT MEFs stimulated with poly(I:C) for 9 hours (PIC-stimulated). The data shown are from one of two ( f and g ) or three ( b-d . All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: ** P

    Techniques Used: Expressing, Microarray, Quantitative RT-PCR, Transfection, Western Blot, Cell Culture, Two Tailed Test

    Additional data supporting mtDNA stress-mediated ISG induction in MEFs. a, Western blot probing STAT1, p-STAT1 (Y701), γH2A.X (DNA damage marker) and GAPDH (loading control) in WT MEFs treated with the indicated doses of doxorubicin (Dox) for 24 hours. (n=3 independent experiments). b, qRT-PCR analysis of the indicated ISGs in WT ( Stat1 +/+ ) and Stat1 null ( Stat1 −/− ) littermate MEFs challenged with (+Dox) or without 500 nM Dox for 24 hours. c, WT MEFs treated with (Dox) or without (Ctrl) 1μM doxorubicin were analysed by immunofluorescence for mitochondrial and mtDNA nucleoid morphology by immunofluorescence against HSP60 (Mito., magenta) and DNA (DNA, cyan). (n=3 independent experiments). Images are Z-stack projections and scale bar represents 10 μm. d and f , mtDNA abundance (relative mtDNA copy number) by qPCR with D-loop primers in WT MEFs exposed to (d) 100 μM 2’−3’-dideoxycytidine (ddC) or (f) 450 ng/mL ethidium bromide (EtBr) for 10–12 days. e, g, qRT-PCR analysis of the ISGs Ifit1 and Ifit3 in (e) Control (Ctrl) and ddC-treated MEFs (described in d) challenged with (+Dox) or without (-Dox) 500nM for doxorubicin for 16 hours, and (g) Control (Ctrl) and EtBr-treated MEFs (described in f ) transfected with 2 μg dsDNA90 or lipofectamine only (Mock) for 9 hours. h , mtDNA abundance (relative mtDNA copy number) by qPCR with D-loop primers in LMTK- cells with (ρ+) or without (ρ˚) mtDNA. i , qRT-PCR analysis of the ISGs Ifit1 and Ifit3 in LMTK- ρ+ and ρ˚ cells (described in h) challenged with 500 nM Dox for 24 hours. j and k, MEFs were pre-treated with 100 μM chloramphenicol (Chlo) for 24 hours followed by 500 nM Doxorubicin (Dox) for 24 hours. j, Western blot using an OXPHOS complex cocktail (mtDNA-encoded subunit is in red, nucleus-encoded subunits in black), γH2A.X (DNA damage marker) or actin (loading control) (n=3 independent experiments). k, qRT-PCR analysis of the ISGs Ifit1 and Ifit3. For e and g the data shown are from one of two biological replicates with the error bars indicating the mean ± SD of three technical replicates. For b, d, f, h and i, . All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: ** P
    Figure Legend Snippet: Additional data supporting mtDNA stress-mediated ISG induction in MEFs. a, Western blot probing STAT1, p-STAT1 (Y701), γH2A.X (DNA damage marker) and GAPDH (loading control) in WT MEFs treated with the indicated doses of doxorubicin (Dox) for 24 hours. (n=3 independent experiments). b, qRT-PCR analysis of the indicated ISGs in WT ( Stat1 +/+ ) and Stat1 null ( Stat1 −/− ) littermate MEFs challenged with (+Dox) or without 500 nM Dox for 24 hours. c, WT MEFs treated with (Dox) or without (Ctrl) 1μM doxorubicin were analysed by immunofluorescence for mitochondrial and mtDNA nucleoid morphology by immunofluorescence against HSP60 (Mito., magenta) and DNA (DNA, cyan). (n=3 independent experiments). Images are Z-stack projections and scale bar represents 10 μm. d and f , mtDNA abundance (relative mtDNA copy number) by qPCR with D-loop primers in WT MEFs exposed to (d) 100 μM 2’−3’-dideoxycytidine (ddC) or (f) 450 ng/mL ethidium bromide (EtBr) for 10–12 days. e, g, qRT-PCR analysis of the ISGs Ifit1 and Ifit3 in (e) Control (Ctrl) and ddC-treated MEFs (described in d) challenged with (+Dox) or without (-Dox) 500nM for doxorubicin for 16 hours, and (g) Control (Ctrl) and EtBr-treated MEFs (described in f ) transfected with 2 μg dsDNA90 or lipofectamine only (Mock) for 9 hours. h , mtDNA abundance (relative mtDNA copy number) by qPCR with D-loop primers in LMTK- cells with (ρ+) or without (ρ˚) mtDNA. i , qRT-PCR analysis of the ISGs Ifit1 and Ifit3 in LMTK- ρ+ and ρ˚ cells (described in h) challenged with 500 nM Dox for 24 hours. j and k, MEFs were pre-treated with 100 μM chloramphenicol (Chlo) for 24 hours followed by 500 nM Doxorubicin (Dox) for 24 hours. j, Western blot using an OXPHOS complex cocktail (mtDNA-encoded subunit is in red, nucleus-encoded subunits in black), γH2A.X (DNA damage marker) or actin (loading control) (n=3 independent experiments). k, qRT-PCR analysis of the ISGs Ifit1 and Ifit3. For e and g the data shown are from one of two biological replicates with the error bars indicating the mean ± SD of three technical replicates. For b, d, f, h and i, . All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: ** P

    Techniques Used: Western Blot, Marker, Quantitative RT-PCR, Immunofluorescence, Real-time Polymerase Chain Reaction, Transfection, Two Tailed Test

    Related Articles

    MTT Assay:

    Article Title: Highlight article: Efficacy of radiation exposure in laryngeal squamous cell carcinoma is mediated by the LAMP3/LAMC2/tenascin-C pathway
    Article Snippet: .. Lipofectamine 2000 reagent was used for transfection of LAMP3-silencing and LAMP-overexpression plasmids, and cell viability was assessed using an MTT assay at 48 h after transfection. .. Each well was supplemented with 10 µL of 5 mg/mL MTT (Sigma-Aldrich, St. Louis, USA) and incubated for 4 h; the MTT solution was removed, and dimethyl sulfoxide was added to each well to dissolve the metabolic product.

    Transfection:

    Article Title: Highlight article: Efficacy of radiation exposure in laryngeal squamous cell carcinoma is mediated by the LAMP3/LAMC2/tenascin-C pathway
    Article Snippet: .. Lipofectamine 2000 reagent was used for transfection of LAMP3-silencing and LAMP-overexpression plasmids, and cell viability was assessed using an MTT assay at 48 h after transfection. .. Each well was supplemented with 10 µL of 5 mg/mL MTT (Sigma-Aldrich, St. Louis, USA) and incubated for 4 h; the MTT solution was removed, and dimethyl sulfoxide was added to each well to dissolve the metabolic product.

    Article Title: Integrated role of microRNA-30e-5p through targeting negative regulators of innate immune pathways during HBV infection and SLE
    Article Snippet: .. Reagents used for transfection/electroporation were Lipofectamine 2000, miRNA mimics, miRNA inhibitors, controls mimic/inhibitors and for RNA isolation were Trizol/Trizol-LS. ..

    Article Title: Mitochondrial DNA Stress Signalling Protects the Nuclear Genome
    Article Snippet: .. Transfection of poly I:C and dsDNA-90 were performed using Lipofectamine 2000 at a ratio of 2:1 (lipofectamine 2000:μg nucleic acid). .. Transfection of siRNAs into MEFs was achieved using Lipofectamine RNAiMax according to manufacturer’s instructions.

    Article Title: P2Y14 receptor has a critical role in acute gouty arthritis by regulating pyroptosis of macrophages
    Article Snippet: .. 3’–5’: AUUCCGACUUGACUUAAGGTT) was used for transient transfection in the presence of lipofectamine 2000. .. Forskolin and SQ22536 were respectively dissolved in DMSO at the concentration of 10 mm as stock solutions.

    Luciferase:

    Article Title: MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
    Article Snippet: .. For the luciferase assay in SOSP-9607 cells, cells at the density of 1.2×105 per well in 24-well plates were cotransfected with 0.8 ug pMIR-REPORT luciferase reporters with 3′-UTR or mut-3′-UTR of PTEN, 100 nM miR-221 mimic or scramble oligonucleotide using Lipofectamine 2000 reagent. .. The cells were also transfected with 50 ng pRL-TK vector as an internal standard.

    Isolation:

    Article Title: Integrated role of microRNA-30e-5p through targeting negative regulators of innate immune pathways during HBV infection and SLE
    Article Snippet: .. Reagents used for transfection/electroporation were Lipofectamine 2000, miRNA mimics, miRNA inhibitors, controls mimic/inhibitors and for RNA isolation were Trizol/Trizol-LS. ..

    Cell Culture:

    Article Title: Biodegradable Polymeric Nanoparticles Show High Efficacy and Specificity at DNA Delivery to Human Glioblastoma in Vitro and in Vivo
    Article Snippet: .. Materials Lipofectamine 2000 (Invitrogen, Carlsbad, CA), Opti-MEM I (Invitrogen), pEGFP-N1 DNA (Elim Biopharmaceuticals, Hayward, CA), pDsRed-Max-N1 (Addgene DNA plasmid 21718, Cambridge, MA), and cell culture media components were used as received. .. Monomers used for synthesizing polymers ( ) were purchased as follows: 1,3-butanediol diacrylate (B3b; Sigma-Aldrich, St. Louis, MO); 1,4-butanediol diacrylate (B4; Alfa Aesar, Ward Hill, MA); 1,5-pentanediol diacrylate (B5; Monomer-Polymer and Dajac Laboratories, Trevose, PA); 3-amino-1-propanol (S3; Alfa Aesar); 4-amino-1-butanol (S4; Alfa Aesar); 5-amino-1-pentanol (S5; Alfa Aesar); 1,3-diaminopropane (E1; Sigma-Aldrich); 1,3-diaminopentane (E3; TCI America, Portland, OR); 2-methyl-1,5-diaminopentane (E4; TCI America); 1,11-diamino-3,6,9-trioxaundecane (E5; TCI America); 2-(3-aminopropylamino)ethanol (E6; Sigma-Aldrich); 1-(3-aminopropyl)-4-methylpiperazine (E7; Alfa Aesar); 1-(3-aminopropyl)pyrrolidine (E8; TCI America); and cystamine dihydrochloride (E10; Alfa Aesar).

    other:

    Article Title: Polymorphic α-Synuclein Strains Modified by Dopamine and Docosahexaenoic Acid Interact Differentially with Tau Protein
    Article Snippet: Cells were exposed to two different concentrations of each tau aggregate strain (0.25 and 0.5 μM) in presence of Lipofectamine 2000 for 24 h followed by washing 3 times with PBS.

    Plasmid Preparation:

    Article Title: Biodegradable Polymeric Nanoparticles Show High Efficacy and Specificity at DNA Delivery to Human Glioblastoma in Vitro and in Vivo
    Article Snippet: .. Materials Lipofectamine 2000 (Invitrogen, Carlsbad, CA), Opti-MEM I (Invitrogen), pEGFP-N1 DNA (Elim Biopharmaceuticals, Hayward, CA), pDsRed-Max-N1 (Addgene DNA plasmid 21718, Cambridge, MA), and cell culture media components were used as received. .. Monomers used for synthesizing polymers ( ) were purchased as follows: 1,3-butanediol diacrylate (B3b; Sigma-Aldrich, St. Louis, MO); 1,4-butanediol diacrylate (B4; Alfa Aesar, Ward Hill, MA); 1,5-pentanediol diacrylate (B5; Monomer-Polymer and Dajac Laboratories, Trevose, PA); 3-amino-1-propanol (S3; Alfa Aesar); 4-amino-1-butanol (S4; Alfa Aesar); 5-amino-1-pentanol (S5; Alfa Aesar); 1,3-diaminopropane (E1; Sigma-Aldrich); 1,3-diaminopentane (E3; TCI America, Portland, OR); 2-methyl-1,5-diaminopentane (E4; TCI America); 1,11-diamino-3,6,9-trioxaundecane (E5; TCI America); 2-(3-aminopropylamino)ethanol (E6; Sigma-Aldrich); 1-(3-aminopropyl)-4-methylpiperazine (E7; Alfa Aesar); 1-(3-aminopropyl)pyrrolidine (E8; TCI America); and cystamine dihydrochloride (E10; Alfa Aesar).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher lipofectamine 2000
    miR-221 targets PTEN leading to activation of the Akt pathway. A , putative miR-221-binding sites (red) in the 3′-UTRs of PTEN ( top ). The middle panel shows sequence alignment of the miR-221 sequences (red color) with a region of the PTEN 3′-UTR (red) from indicated species. The bottom shows a mutant of PTEN 3′-UTR for pMIR-REPORT. B , SOSP-9607 cells were transfected with miR-221 mimic, miR-221 inhibitor, scramble oligonucleotide or nothing (blank). And then indicated protein and mRNA were detected. Rows 1 to 7, the expression of PTEN, phospho-Akt (p-Akt), total-Akt (t-Akt), Cyclin D1 (CCND1), p27 and p57 were analyzed by Western blot; row 8, β-actin was used as a loading control; row 9, PTEN mRNA level was measured by RT-PCR; row 10, GAPDH was used as an control. C , SOSP-9607 cells were co-transfected with miR-221 mimic or scramble oligonucleotide, and pMIR-REPORT vector consisting of a luciferase gene containing the wild type (PTEN-3′-UTR) or mutated (PTEN-mut-3′-UTR) miR-221 binding sites in its 3′-UTR region by using <t>Lipofectamine</t> 2000 reagent. The cells were also transfected with 50 ng pRL-TK vector as an internal standard. 24 h after transfection, luciferase activity was measured by using a dual luciferase reporter assay (Promega). D , similar experiments were performed in MG63 cells as described in panel C except that miR-221 mimic was instead by miR-221 inhibitor. The results were presented as relative luciferase activity (firefly Luc/Renilla Luc). All experiments were repeated three times in triplicate. Columns, mean of three independent experiments; bars, SD. **, p
    Lipofectamine 2000, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 75677 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipofectamine 2000/product/Thermo Fisher
    Average 99 stars, based on 75677 article reviews
    Price from $9.99 to $1999.99
    lipofectamine 2000 - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    Image Search Results


    miR-221 targets PTEN leading to activation of the Akt pathway. A , putative miR-221-binding sites (red) in the 3′-UTRs of PTEN ( top ). The middle panel shows sequence alignment of the miR-221 sequences (red color) with a region of the PTEN 3′-UTR (red) from indicated species. The bottom shows a mutant of PTEN 3′-UTR for pMIR-REPORT. B , SOSP-9607 cells were transfected with miR-221 mimic, miR-221 inhibitor, scramble oligonucleotide or nothing (blank). And then indicated protein and mRNA were detected. Rows 1 to 7, the expression of PTEN, phospho-Akt (p-Akt), total-Akt (t-Akt), Cyclin D1 (CCND1), p27 and p57 were analyzed by Western blot; row 8, β-actin was used as a loading control; row 9, PTEN mRNA level was measured by RT-PCR; row 10, GAPDH was used as an control. C , SOSP-9607 cells were co-transfected with miR-221 mimic or scramble oligonucleotide, and pMIR-REPORT vector consisting of a luciferase gene containing the wild type (PTEN-3′-UTR) or mutated (PTEN-mut-3′-UTR) miR-221 binding sites in its 3′-UTR region by using Lipofectamine 2000 reagent. The cells were also transfected with 50 ng pRL-TK vector as an internal standard. 24 h after transfection, luciferase activity was measured by using a dual luciferase reporter assay (Promega). D , similar experiments were performed in MG63 cells as described in panel C except that miR-221 mimic was instead by miR-221 inhibitor. The results were presented as relative luciferase activity (firefly Luc/Renilla Luc). All experiments were repeated three times in triplicate. Columns, mean of three independent experiments; bars, SD. **, p

    Journal: PLoS ONE

    Article Title: MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma

    doi: 10.1371/journal.pone.0053906

    Figure Lengend Snippet: miR-221 targets PTEN leading to activation of the Akt pathway. A , putative miR-221-binding sites (red) in the 3′-UTRs of PTEN ( top ). The middle panel shows sequence alignment of the miR-221 sequences (red color) with a region of the PTEN 3′-UTR (red) from indicated species. The bottom shows a mutant of PTEN 3′-UTR for pMIR-REPORT. B , SOSP-9607 cells were transfected with miR-221 mimic, miR-221 inhibitor, scramble oligonucleotide or nothing (blank). And then indicated protein and mRNA were detected. Rows 1 to 7, the expression of PTEN, phospho-Akt (p-Akt), total-Akt (t-Akt), Cyclin D1 (CCND1), p27 and p57 were analyzed by Western blot; row 8, β-actin was used as a loading control; row 9, PTEN mRNA level was measured by RT-PCR; row 10, GAPDH was used as an control. C , SOSP-9607 cells were co-transfected with miR-221 mimic or scramble oligonucleotide, and pMIR-REPORT vector consisting of a luciferase gene containing the wild type (PTEN-3′-UTR) or mutated (PTEN-mut-3′-UTR) miR-221 binding sites in its 3′-UTR region by using Lipofectamine 2000 reagent. The cells were also transfected with 50 ng pRL-TK vector as an internal standard. 24 h after transfection, luciferase activity was measured by using a dual luciferase reporter assay (Promega). D , similar experiments were performed in MG63 cells as described in panel C except that miR-221 mimic was instead by miR-221 inhibitor. The results were presented as relative luciferase activity (firefly Luc/Renilla Luc). All experiments were repeated three times in triplicate. Columns, mean of three independent experiments; bars, SD. **, p

    Article Snippet: For the luciferase assay in SOSP-9607 cells, cells at the density of 1.2×105 per well in 24-well plates were cotransfected with 0.8 ug pMIR-REPORT luciferase reporters with 3′-UTR or mut-3′-UTR of PTEN, 100 nM miR-221 mimic or scramble oligonucleotide using Lipofectamine 2000 reagent.

    Techniques: Activation Assay, Binding Assay, Sequencing, Mutagenesis, Transfection, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Luciferase, Activity Assay, Reporter Assay

    Seeding assay of tau aggregate strains in Tau RD-P301S CFP/YFP FRET biosensor cells. a – d Tau biosensor cells were exposed to the cross-seeded tau aggregates or unseeded tau aggregate at 0.25 and 0.5 μM concentrations for 24 h with Lipofectamine. The cross-seeded tau aggregates, TauO SynO-DA and TauO SynO-DHA show dose-dependent increased seeding propensity at the concentrations used resulting in the formation of cytosolic tau aggregates. Unseeded tau aggregates do not show any seeding. TauO SynO-DA has more potent seeding efficiency than TauO SynO-DHA . e Quantification of FRET positive cells observed in all 4 groups of treatment: vehicle, TauO unseeded , TauO SynO-DA , and TauO SynO-DHA . Quantification is performed from fifteen regions of interest (ROIs) from three replicates performed in three independent experiments. The histograms represent mean ± SD. Statistical significance is measured by using two-way ANOVA with Bonferroni post hoc analysis. ** p

    Journal: Molecular Neurobiology

    Article Title: Polymorphic α-Synuclein Strains Modified by Dopamine and Docosahexaenoic Acid Interact Differentially with Tau Protein

    doi: 10.1007/s12035-020-01913-6

    Figure Lengend Snippet: Seeding assay of tau aggregate strains in Tau RD-P301S CFP/YFP FRET biosensor cells. a – d Tau biosensor cells were exposed to the cross-seeded tau aggregates or unseeded tau aggregate at 0.25 and 0.5 μM concentrations for 24 h with Lipofectamine. The cross-seeded tau aggregates, TauO SynO-DA and TauO SynO-DHA show dose-dependent increased seeding propensity at the concentrations used resulting in the formation of cytosolic tau aggregates. Unseeded tau aggregates do not show any seeding. TauO SynO-DA has more potent seeding efficiency than TauO SynO-DHA . e Quantification of FRET positive cells observed in all 4 groups of treatment: vehicle, TauO unseeded , TauO SynO-DA , and TauO SynO-DHA . Quantification is performed from fifteen regions of interest (ROIs) from three replicates performed in three independent experiments. The histograms represent mean ± SD. Statistical significance is measured by using two-way ANOVA with Bonferroni post hoc analysis. ** p

    Article Snippet: Cells were exposed to two different concentrations of each tau aggregate strain (0.25 and 0.5 μM) in presence of Lipofectamine 2000 for 24 h followed by washing 3 times with PBS.

    Techniques:

    Human PBMCs stimulated with DAMPs induce miRNA-30e and enhance innate immune responses: PBMCs from three healthy individuals were transfected with their own genomic DNA (ds DNA). (A) Isolated genomic DNA sonicated into small fragments of dsDNA of approximately 100-150 bps each (as shown) and transfected using Lipofectamine 2000. (B-F) Quantification (by qRT-PCR analysis) of the fold changes in the relative abundances of (B) miR-30e, (C-E) respective innate immune transcripts ( IFNα, IFNβ, IFIT1  and  IL6)  in all individuals and (F) NDV viral transcript (shown as per schematic workflow). Data are mean +/-SEM of triplicate samples from single experiment and are representative of three independent experiments in three different individuals. *** P

    Journal: bioRxiv

    Article Title: Integrated role of microRNA-30e-5p through targeting negative regulators of innate immune pathways during HBV infection and SLE

    doi: 10.1101/2020.02.29.969014

    Figure Lengend Snippet: Human PBMCs stimulated with DAMPs induce miRNA-30e and enhance innate immune responses: PBMCs from three healthy individuals were transfected with their own genomic DNA (ds DNA). (A) Isolated genomic DNA sonicated into small fragments of dsDNA of approximately 100-150 bps each (as shown) and transfected using Lipofectamine 2000. (B-F) Quantification (by qRT-PCR analysis) of the fold changes in the relative abundances of (B) miR-30e, (C-E) respective innate immune transcripts ( IFNα, IFNβ, IFIT1 and IL6) in all individuals and (F) NDV viral transcript (shown as per schematic workflow). Data are mean +/-SEM of triplicate samples from single experiment and are representative of three independent experiments in three different individuals. *** P

    Article Snippet: Reagents used for transfection/electroporation were Lipofectamine 2000, miRNA mimics, miRNA inhibitors, controls mimic/inhibitors and for RNA isolation were Trizol/Trizol-LS.

    Techniques: Transfection, Isolation, Sonication, Quantitative RT-PCR

    The effect of LAMP3 on cell viability and apoptosis in HEp-2 cells. (a) Measurements of cell viability from cultured HEp-2 cells after LAMP3 knockdown or overexpression. HEp-2 cells were cultured in 96-well plates. LAMP3 knockdown and overexpression plasmids were transfected into cells using Lipofectamine 2000. After 48 h, the proliferation of HEp-2 cells was measured by the MTT assay. (b, c and d) Apoptosis in HEp-2 cells was assessed using the Annexin V-FITC/PI Apoptosis Detection kit, and fluorescence-activated cell sorting analysis was performed. The upper right quadrant (Q2; Annexin V + /PI + ) indicates apoptosis. Cell proliferation was significantly inhibited and apoptosis increased after LAMP3 knockdown; however, the inhibitory effect was attenuated and apoptosis decreased after LAMP3 complementation. The experiment was performed in triplicate, and representative data are shown. ** P

    Journal: Experimental Biology and Medicine

    Article Title: Highlight article: Efficacy of radiation exposure in laryngeal squamous cell carcinoma is mediated by the LAMP3/LAMC2/tenascin-C pathway

    doi: 10.1177/1535370219867643

    Figure Lengend Snippet: The effect of LAMP3 on cell viability and apoptosis in HEp-2 cells. (a) Measurements of cell viability from cultured HEp-2 cells after LAMP3 knockdown or overexpression. HEp-2 cells were cultured in 96-well plates. LAMP3 knockdown and overexpression plasmids were transfected into cells using Lipofectamine 2000. After 48 h, the proliferation of HEp-2 cells was measured by the MTT assay. (b, c and d) Apoptosis in HEp-2 cells was assessed using the Annexin V-FITC/PI Apoptosis Detection kit, and fluorescence-activated cell sorting analysis was performed. The upper right quadrant (Q2; Annexin V + /PI + ) indicates apoptosis. Cell proliferation was significantly inhibited and apoptosis increased after LAMP3 knockdown; however, the inhibitory effect was attenuated and apoptosis decreased after LAMP3 complementation. The experiment was performed in triplicate, and representative data are shown. ** P

    Article Snippet: Lipofectamine 2000 reagent was used for transfection of LAMP3-silencing and LAMP-overexpression plasmids, and cell viability was assessed using an MTT assay at 48 h after transfection.

    Techniques: Cell Culture, Over Expression, Transfection, MTT Assay, Fluorescence, FACS