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    Millipore g 418 sulfate
    The effect of miR-29b on mESCs differentiation. ( A ) Scheme of the constructs used to express miRNA of interest. Representative bright-field images from pcDNA-pre-miR-29b mESCs after five or eight days of <t>G418</t> induction. Scale bars: 50 μm. ( B ) The effect of miR-29b on pluripotent, ectoderm and mesendoderm markers expression in mESCs by qPCR. ( C ) Expression of miR-29b precusor in 15 mouse tissues was measured by qRT-PCR and normalized to Gapdh mRNA levels. Each bar in the figure represents the mean ± SEM of triplicates.
    G 418 Sulfate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 228 article reviews
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    99
    Millipore g418
    smFRET experiments. (A) eEF2-induced translocation of the 80S-Trp-IRES-PRE6 complex to form 80S-Trp-IRES-POST6 complex followed by release of tRNA Gln . The cartoon at the top shows the state progression during translocation. (i) Single molecule traces. Green and red traces show tRNA Trp (Cy3) emission and tRNA Gln (Cy5) sensitized emission, respectively, following eEF2 injection, excited at 532 nm. (ii) ALEX intensity signal from direct excitation of tRNA Gln (Cy5) at 640 nm. (iii) FRET efficiency between tRNA Trp (Cy3) and tRNA Gln (Cy5) showing a transient increase following eEF2 injection on conversion of the PRE6 complex to POST6. (B) Recording of fluorescence traces for a Stop-IRES complex that did not bind tRNA Trp . Colors as in panel A. (C) Effects of increasing <t>G418</t> concentration on PRE6 complex formation from 80S-Stop-IRES-POST5 complex. Added G418 did not affect formation of Trp-IRES-PRE6 from Trp-IRES-POST5.
    G418, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 20741 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/g418/product/Millipore
    Average 99 stars, based on 20741 article reviews
    Price from $9.99 to $1999.99
    g418 - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    The effect of miR-29b on mESCs differentiation. ( A ) Scheme of the constructs used to express miRNA of interest. Representative bright-field images from pcDNA-pre-miR-29b mESCs after five or eight days of G418 induction. Scale bars: 50 μm. ( B ) The effect of miR-29b on pluripotent, ectoderm and mesendoderm markers expression in mESCs by qPCR. ( C ) Expression of miR-29b precusor in 15 mouse tissues was measured by qRT-PCR and normalized to Gapdh mRNA levels. Each bar in the figure represents the mean ± SEM of triplicates.

    Journal: Nucleic Acids Research

    Article Title: MicroRNA-29b/Tet1 regulatory axis epigenetically modulates mesendoderm differentiation in mouse embryonic stem cells

    doi: 10.1093/nar/gkv653

    Figure Lengend Snippet: The effect of miR-29b on mESCs differentiation. ( A ) Scheme of the constructs used to express miRNA of interest. Representative bright-field images from pcDNA-pre-miR-29b mESCs after five or eight days of G418 induction. Scale bars: 50 μm. ( B ) The effect of miR-29b on pluripotent, ectoderm and mesendoderm markers expression in mESCs by qPCR. ( C ) Expression of miR-29b precusor in 15 mouse tissues was measured by qRT-PCR and normalized to Gapdh mRNA levels. Each bar in the figure represents the mean ± SEM of triplicates.

    Article Snippet: G418 selection (1 mg/ml, Sigma) was conducted for 2 weeks to select the miR-29b stable overexpressing mESCs.

    Techniques: Construct, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Generation of biPSCs. (A) Maps of plasmid vectors pSTEM-h103 and pCAG-PBase. The pSTEM-h103 contains eight reprogramming factors, PB sequences, and drug screening genes Neo and TK. The pCAG-PBase contains CAG-promoted PBase that can effect transposition of PB sequences into cell genome. (B) Schematic diagram of the reprogramming protocols used in the experiment. Bat embryonic fibroblasts isolated from bats of Myotis lucifugus were transfected with pSTEM-h103 and pCAG-PBase simultaneously and cultured on feeder layers with MEF medium. After 1 day, the medium was changed into ESC medium with G418. On Day 7, the medium was replaced with 3i medium, which was beneficial for the generation of iPSCs of high quality. The colonies could be picked from Day 14. (C) Bat embryonic fibroblasts derived from little brown bats of Myotis lucifugus (left). A representative biPSC colony with mouse ESC-like morphology at passage 45 is shown at a low magnification (middle) and at a higher magnification (right). biPSC, bat-induced pluripotent stem cell; MEF, mouse embryonic fibroblast; ESC, embryonic stem cell (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).

    Journal: Theriogenology

    Article Title: Generation and characterization of bat-induced pluripotent stem cells

    doi: 10.1016/j.theriogenology.2014.04.001

    Figure Lengend Snippet: Generation of biPSCs. (A) Maps of plasmid vectors pSTEM-h103 and pCAG-PBase. The pSTEM-h103 contains eight reprogramming factors, PB sequences, and drug screening genes Neo and TK. The pCAG-PBase contains CAG-promoted PBase that can effect transposition of PB sequences into cell genome. (B) Schematic diagram of the reprogramming protocols used in the experiment. Bat embryonic fibroblasts isolated from bats of Myotis lucifugus were transfected with pSTEM-h103 and pCAG-PBase simultaneously and cultured on feeder layers with MEF medium. After 1 day, the medium was changed into ESC medium with G418. On Day 7, the medium was replaced with 3i medium, which was beneficial for the generation of iPSCs of high quality. The colonies could be picked from Day 14. (C) Bat embryonic fibroblasts derived from little brown bats of Myotis lucifugus (left). A representative biPSC colony with mouse ESC-like morphology at passage 45 is shown at a low magnification (middle) and at a higher magnification (right). biPSC, bat-induced pluripotent stem cell; MEF, mouse embryonic fibroblast; ESC, embryonic stem cell (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).

    Article Snippet: The concentration of G418 sulfate (Calbiochem) for BEFs after nucleofection was 500 μg/mL.

    Techniques: Plasmid Preparation, Isolation, Transfection, Cell Culture, Derivative Assay

    Reconstruction of bacterial populations in D. melanogaster following toxic treatment. a Naïve flies using six regions (left) and one region (right). b Same for flies reared on medium containing the G418 toxin. c Ambiguity of the three most abundant species. Profiling based on a single region (hollow bars) and six regions (full bars). d Similarity between L. plantarum predicted strains and the Sanger sequence of the strain isolated from the detected colony (top 100 sequences)

    Journal: Microbiome

    Article Title: Combining 16S rRNA gene variable regions enables high-resolution microbial community profiling

    doi: 10.1186/s40168-017-0396-x

    Figure Lengend Snippet: Reconstruction of bacterial populations in D. melanogaster following toxic treatment. a Naïve flies using six regions (left) and one region (right). b Same for flies reared on medium containing the G418 toxin. c Ambiguity of the three most abundant species. Profiling based on a single region (hollow bars) and six regions (full bars). d Similarity between L. plantarum predicted strains and the Sanger sequence of the strain isolated from the detected colony (top 100 sequences)

    Article Snippet: Extracting bacterial DNA Flies were reared either on standard medium or medium containing 400 μg/ml of the G418 toxin (Sigma).

    Techniques: Sequencing, Isolation

    Generation and verification of tagged protein arrays. ( A ) To tag ORFX as bait (V5–6×HIS) and prey (V5–3×VSV), a set of primers is used that anneal to identical binding sites within the template plasmids and have flanking sequence homologous to ORFX . PCR products generated from the bait and prey templates are transformed into a - and α-cells, respectively. Homologous recombination occurs between the variable portion of the 5′ primer (light blue) and the 3′ terminus of the ORF, and between the variable portion of the 3′ primer (red) and the 3′ UTR) of ORFX . Transformants are selected on G418 plates, and colony PCR is performed to verify integration of the Kan r downstream of the desired ORF. Abbreviations: TEF, translational elongation factor; TEFp, TEF promoter; TEFt, TEF terminator: Kan r , kanamycin resistance; loxp, site for CRE specific homologous recombination. ( B . The asterisk (*) denotes possible misloading or protein degradation. Note in the RAD51 lane the multiple protein products. Expected protein sizes are listed in Supplemental Table 1. ( C ) Analysis of effects on cell growth by tagging essential genes. A total of 24 strains with essential genes tagged as baits (6×HIS) and preys (3×VSV) were grown to saturation and spotted in 10-fold dilutions on YPD. Pictures were taken after 2 d at 30°C.

    Journal: Genome Research

    Article Title: Examining protein-protein interactions using endogenously tagged yeast arrays: The Cross-and-Capture system

    doi: 10.1101/gr.6667007

    Figure Lengend Snippet: Generation and verification of tagged protein arrays. ( A ) To tag ORFX as bait (V5–6×HIS) and prey (V5–3×VSV), a set of primers is used that anneal to identical binding sites within the template plasmids and have flanking sequence homologous to ORFX . PCR products generated from the bait and prey templates are transformed into a - and α-cells, respectively. Homologous recombination occurs between the variable portion of the 5′ primer (light blue) and the 3′ terminus of the ORF, and between the variable portion of the 3′ primer (red) and the 3′ UTR) of ORFX . Transformants are selected on G418 plates, and colony PCR is performed to verify integration of the Kan r downstream of the desired ORF. Abbreviations: TEF, translational elongation factor; TEFp, TEF promoter; TEFt, TEF terminator: Kan r , kanamycin resistance; loxp, site for CRE specific homologous recombination. ( B . The asterisk (*) denotes possible misloading or protein degradation. Note in the RAD51 lane the multiple protein products. Expected protein sizes are listed in Supplemental Table 1. ( C ) Analysis of effects on cell growth by tagging essential genes. A total of 24 strains with essential genes tagged as baits (6×HIS) and preys (3×VSV) were grown to saturation and spotted in 10-fold dilutions on YPD. Pictures were taken after 2 d at 30°C.

    Article Snippet: Transformants that grew on the G418 plates were restreaked and tested for proper integration of the tagging cassette via colony PCR.

    Techniques: Binding Assay, Sequencing, Polymerase Chain Reaction, Generated, Transformation Assay, Homologous Recombination

    Schematic representation of plasmid vectors used for stable transformation of Giardia trophozoites. A RAN-neo cassette conferring constitutive G418 resistance is cloned immediately adjacent to a recombinant CWP1 locus in a head-to-head conformation on a pBluescript backbone. All four domains of CWP1 (I–IV) are sequentially added to the N-terminal side of a GFP reporter gene. In construct pC0-PP2, the CWP1 3′ flanking sequence was replaced by that of the Giardia PP2 gene. CWP1 domains are indicated by roman numerals. Promoters and corresponding direction of transcription are represented by bent arrows.

    Journal: Molecular Biology of the Cell

    Article Title: Stage-Specific Expression and Targeting of Cyst Wall Protein-Green Fluorescent Protein Chimeras in Giardia

    doi:

    Figure Lengend Snippet: Schematic representation of plasmid vectors used for stable transformation of Giardia trophozoites. A RAN-neo cassette conferring constitutive G418 resistance is cloned immediately adjacent to a recombinant CWP1 locus in a head-to-head conformation on a pBluescript backbone. All four domains of CWP1 (I–IV) are sequentially added to the N-terminal side of a GFP reporter gene. In construct pC0-PP2, the CWP1 3′ flanking sequence was replaced by that of the Giardia PP2 gene. CWP1 domains are indicated by roman numerals. Promoters and corresponding direction of transcription are represented by bent arrows.

    Article Snippet: For selection of stable transformants the parasites were maintained in TYI-S-33 culture medium containing the antibiotic G418 (Sigma, St. Louis, MO) at a concentration of 135 μg/ml.

    Techniques: Plasmid Preparation, Transformation Assay, Clone Assay, Recombinant, Construct, Sequencing

    pAc5-STABLE1-Neo confers G418 resistance to S2R+ cells. (A–I) Bright field images of S2R+ Drosophila cells that were mock-transfected (Ctrl; A–C), transfected with pAc5-GFP (D–F) or with pAc5-STABLE1-Neo (G–I). Three days after transfection, G418 was added to the media at concentrations of 0 (A, D, G), 600 (B, E, H) or 2000 μg/ml (C, F, I). (A'–I') Fluorescent images of the same cells from panels A to I showing GFP expression. (J–L) Percentage of surviving cells that are positive for GFP. Letters correspond to panels/treatments directly above each graph. All images were taken after 30 days of treatment with G418.

    Journal: Scientific Reports

    Article Title: Generation of stable Drosophila cell lines using multicistronic vectors

    doi: 10.1038/srep00075

    Figure Lengend Snippet: pAc5-STABLE1-Neo confers G418 resistance to S2R+ cells. (A–I) Bright field images of S2R+ Drosophila cells that were mock-transfected (Ctrl; A–C), transfected with pAc5-GFP (D–F) or with pAc5-STABLE1-Neo (G–I). Three days after transfection, G418 was added to the media at concentrations of 0 (A, D, G), 600 (B, E, H) or 2000 μg/ml (C, F, I). (A'–I') Fluorescent images of the same cells from panels A to I showing GFP expression. (J–L) Percentage of surviving cells that are positive for GFP. Letters correspond to panels/treatments directly above each graph. All images were taken after 30 days of treatment with G418.

    Article Snippet: After 72 hours, selective medium was added with different concentrations (0, 600, 2000 μg/ml) of neomycin/G418 (Sigma).

    Techniques: Transfection, Expressing

    Expression of two different proteins as well as a selectable marker from a single vector in Drosophila cultured cells. (A–I) Brightfield images of Drosophila S2R+ cells that were mock-transfected (Ctrl; A–C), transfected with pAc5-GFP (D–F) or transfected with pAc5-STABLE2-Neo (G–I). Three days after transfection, G418 was added to the media at concentrations of 0 (A, D, G), 600 (B, E, H) or 2000 μg/ml (C, F, I). Fluorescent images of the same cells from panels A–I showing GFP expression (A'–I') or FLAG-mCherry expression (A”–I”). White boxes mark zoomed region shown in inset (panel I, overlay). All images were taken after 30 days of treatment with G418.

    Journal: Scientific Reports

    Article Title: Generation of stable Drosophila cell lines using multicistronic vectors

    doi: 10.1038/srep00075

    Figure Lengend Snippet: Expression of two different proteins as well as a selectable marker from a single vector in Drosophila cultured cells. (A–I) Brightfield images of Drosophila S2R+ cells that were mock-transfected (Ctrl; A–C), transfected with pAc5-GFP (D–F) or transfected with pAc5-STABLE2-Neo (G–I). Three days after transfection, G418 was added to the media at concentrations of 0 (A, D, G), 600 (B, E, H) or 2000 μg/ml (C, F, I). Fluorescent images of the same cells from panels A–I showing GFP expression (A'–I') or FLAG-mCherry expression (A”–I”). White boxes mark zoomed region shown in inset (panel I, overlay). All images were taken after 30 days of treatment with G418.

    Article Snippet: After 72 hours, selective medium was added with different concentrations (0, 600, 2000 μg/ml) of neomycin/G418 (Sigma).

    Techniques: Expressing, Marker, Plasmid Preparation, Cell Culture, Transfection

    pAc5-STABLE1-Neo confers G418 resistance in Kc167 cells. (A–I) Bright field images of Drosophila Kc167 cells that were mock-transfected (A–C), transfected with pAc5-GFP (D–F) or transfected with pAc5-STABLE1-Neo (G–I). Three days after transfection, G418 was added to the media at concentrations of 0 (A, D, G), 600 (B, E, H) or 2000 μg/ml (C, F, I). (A'–I') Fluorescent images of the same cells from panels A to I showing GFP expression. (J–L) Percentage of cells that are positive for GFP. Letters correspond to panels/treatments directly above each graph. All images were taken after 30 days of treatment with G418.

    Journal: Scientific Reports

    Article Title: Generation of stable Drosophila cell lines using multicistronic vectors

    doi: 10.1038/srep00075

    Figure Lengend Snippet: pAc5-STABLE1-Neo confers G418 resistance in Kc167 cells. (A–I) Bright field images of Drosophila Kc167 cells that were mock-transfected (A–C), transfected with pAc5-GFP (D–F) or transfected with pAc5-STABLE1-Neo (G–I). Three days after transfection, G418 was added to the media at concentrations of 0 (A, D, G), 600 (B, E, H) or 2000 μg/ml (C, F, I). (A'–I') Fluorescent images of the same cells from panels A to I showing GFP expression. (J–L) Percentage of cells that are positive for GFP. Letters correspond to panels/treatments directly above each graph. All images were taken after 30 days of treatment with G418.

    Article Snippet: After 72 hours, selective medium was added with different concentrations (0, 600, 2000 μg/ml) of neomycin/G418 (Sigma).

    Techniques: Transfection, Expressing

    Effect of PKG-Iα expression on high molecular mass pElk-1 in rat aortic SMCs. Rat aortic SMCs at passage 3 to 5 were transfected with control pcDNA3.1 vector (Cont) or PKG-Iα-pcDNA3.1 (PKG-Iα) using Lipofectamine 2000. Stably transfected cells were selected using 500 μg/ml of G418. Cells grown to confluence were serum deprived in DMEM containing 1 mg/ml of BSA for 3 days. A : nuclear extracts or whole cell extracts were prepared and immunoblotted (IB) with indicated antibodies. Arrow on pElk-1 blot indicates high molecular mass pElk-1 *Corresponding high molecular mass Elk-1 signal on Elk-1 blot. Data are a representative of 3 experiments. SM-MHC, smooth muscle-myosin heavy chain; SRF, serum response factor. B : quantitative analysis of 3 pElk-1 blots was performed using ImageJ software. Results are expressed as means ± SD. * P

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: cGMP-dependent protein kinase and the regulation of vascular smooth muscle cell gene expression: possible involvement of Elk-1 sumoylation

    doi: 10.1152/ajpheart.00677.2010

    Figure Lengend Snippet: Effect of PKG-Iα expression on high molecular mass pElk-1 in rat aortic SMCs. Rat aortic SMCs at passage 3 to 5 were transfected with control pcDNA3.1 vector (Cont) or PKG-Iα-pcDNA3.1 (PKG-Iα) using Lipofectamine 2000. Stably transfected cells were selected using 500 μg/ml of G418. Cells grown to confluence were serum deprived in DMEM containing 1 mg/ml of BSA for 3 days. A : nuclear extracts or whole cell extracts were prepared and immunoblotted (IB) with indicated antibodies. Arrow on pElk-1 blot indicates high molecular mass pElk-1 *Corresponding high molecular mass Elk-1 signal on Elk-1 blot. Data are a representative of 3 experiments. SM-MHC, smooth muscle-myosin heavy chain; SRF, serum response factor. B : quantitative analysis of 3 pElk-1 blots was performed using ImageJ software. Results are expressed as means ± SD. * P

    Article Snippet: To acquire stable PKG-transfected rat aortic SMCs, empty or PKG-Iα-expressing plasmids were used for transfection and selected under 500 μg/ml of G418 antibiotics (Sigma) as described previously ( , ).

    Techniques: Expressing, Transfection, Plasmid Preparation, Stable Transfection, Software

    smFRET experiments. (A) eEF2-induced translocation of the 80S-Trp-IRES-PRE6 complex to form 80S-Trp-IRES-POST6 complex followed by release of tRNA Gln . The cartoon at the top shows the state progression during translocation. (i) Single molecule traces. Green and red traces show tRNA Trp (Cy3) emission and tRNA Gln (Cy5) sensitized emission, respectively, following eEF2 injection, excited at 532 nm. (ii) ALEX intensity signal from direct excitation of tRNA Gln (Cy5) at 640 nm. (iii) FRET efficiency between tRNA Trp (Cy3) and tRNA Gln (Cy5) showing a transient increase following eEF2 injection on conversion of the PRE6 complex to POST6. (B) Recording of fluorescence traces for a Stop-IRES complex that did not bind tRNA Trp . Colors as in panel A. (C) Effects of increasing G418 concentration on PRE6 complex formation from 80S-Stop-IRES-POST5 complex. Added G418 did not affect formation of Trp-IRES-PRE6 from Trp-IRES-POST5.

    Journal: ACS Medicinal Chemistry Letters

    Article Title: New in Vitro Assay Measuring Direct Interaction of Nonsense Suppressors with the Eukaryotic Protein Synthesis Machinery

    doi: 10.1021/acsmedchemlett.8b00472

    Figure Lengend Snippet: smFRET experiments. (A) eEF2-induced translocation of the 80S-Trp-IRES-PRE6 complex to form 80S-Trp-IRES-POST6 complex followed by release of tRNA Gln . The cartoon at the top shows the state progression during translocation. (i) Single molecule traces. Green and red traces show tRNA Trp (Cy3) emission and tRNA Gln (Cy5) sensitized emission, respectively, following eEF2 injection, excited at 532 nm. (ii) ALEX intensity signal from direct excitation of tRNA Gln (Cy5) at 640 nm. (iii) FRET efficiency between tRNA Trp (Cy3) and tRNA Gln (Cy5) showing a transient increase following eEF2 injection on conversion of the PRE6 complex to POST6. (B) Recording of fluorescence traces for a Stop-IRES complex that did not bind tRNA Trp . Colors as in panel A. (C) Effects of increasing G418 concentration on PRE6 complex formation from 80S-Stop-IRES-POST5 complex. Added G418 did not affect formation of Trp-IRES-PRE6 from Trp-IRES-POST5.

    Article Snippet: Nonsense suppressors ( ) were obtained as follows: gentamicin mixture and G418 (Sigma), nourseothricin sulfate, a mixture of streptothricins D and F (Gold Biotechnology), doxorubicin (Fisher Scientific), escin and tylosin (Alfa Aesar), azithromycin (APExBIO), gentamicin B (MicroCombiChem).

    Techniques: Translocation Assay, Injection, Fluorescence, Concentration Assay