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  • 96
    Millipore g418
    Functionality of Blast, Puro and <t>G418-resistance</t> genes from pINS plasmids is preserved in the products of recombination . Left part . Colony forming assay. HeLa cells transfected by 1 mkg of indicated plasmids were selected by either blasticidin S, puromycin or G418. After completion of selection the cells were stained by methylene blue. Insertion vectors pINS-Blast, pINS-Neo and pINS-Puro and the products of their recombination with Target vector phrGFP (Blast × GFP, Neo × GFP and Puro × GFP) provide HeLa cells with the resistance to blasticidin S, G418 or puromycin respectively. Right part . Schematic representation of the Cre-mediated recombination between Insertion vectors pINS-Blast, pINS-Neo and pINS-Puro and Target vector phrGFP.
    G418, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 26810 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/g418/product/Millipore
    Average 96 stars, based on 26810 article reviews
    Price from $9.99 to $1999.99
    g418 - by Bioz Stars, 2020-08
    96/100 stars
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    98
    Millipore g418 disulfate salt
    Transient and permanent expression of the transferred DNA. (a) Overview of the experimental design to detect transient expression or stable integration of the transferred DNA. After infection of human cell lines with B. henselae , the DNA transferred through the T4SS will get to the nucleus where genes will be expressed. At 3 days postinfection, transient expression of gfp can be detected by flow cytometry. Antibiotic treatment was applied for long-term selection of neomycin-resistant colonies, to detect stable integration events. (b to d) Graphic representation of the percentage of GFP-positive cells obtained 3 days postinfection (b) and the number of <t>G418-resistant</t> colonies normalized for the number of cells at the beginning of the selection (c), as well as the Neo r /GFP + ratio (d). The different bars represented for each cell line correspond to the different relaxases under study, following the color code indicated in the squares at the top right. Data represent the means of the results from at least 3 independent experiments. Error bars indicate standard deviations. *, P
    G418 Disulfate Salt, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 774 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/g418 disulfate salt/product/Millipore
    Average 98 stars, based on 774 article reviews
    Price from $9.99 to $1999.99
    g418 disulfate salt - by Bioz Stars, 2020-08
    98/100 stars
      Buy from Supplier

    Image Search Results


    Functionality of Blast, Puro and G418-resistance genes from pINS plasmids is preserved in the products of recombination . Left part . Colony forming assay. HeLa cells transfected by 1 mkg of indicated plasmids were selected by either blasticidin S, puromycin or G418. After completion of selection the cells were stained by methylene blue. Insertion vectors pINS-Blast, pINS-Neo and pINS-Puro and the products of their recombination with Target vector phrGFP (Blast × GFP, Neo × GFP and Puro × GFP) provide HeLa cells with the resistance to blasticidin S, G418 or puromycin respectively. Right part . Schematic representation of the Cre-mediated recombination between Insertion vectors pINS-Blast, pINS-Neo and pINS-Puro and Target vector phrGFP.

    Journal: BMC Research Notes

    Article Title: A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination

    doi: 10.1186/1756-0500-1-135

    Figure Lengend Snippet: Functionality of Blast, Puro and G418-resistance genes from pINS plasmids is preserved in the products of recombination . Left part . Colony forming assay. HeLa cells transfected by 1 mkg of indicated plasmids were selected by either blasticidin S, puromycin or G418. After completion of selection the cells were stained by methylene blue. Insertion vectors pINS-Blast, pINS-Neo and pINS-Puro and the products of their recombination with Target vector phrGFP (Blast × GFP, Neo × GFP and Puro × GFP) provide HeLa cells with the resistance to blasticidin S, G418 or puromycin respectively. Right part . Schematic representation of the Cre-mediated recombination between Insertion vectors pINS-Blast, pINS-Neo and pINS-Puro and Target vector phrGFP.

    Article Snippet: 24 hours after transfection media was replaced and the cells were either grown for another 24 hours (transient transfection) or selected with either 3 mg/ml puromycin (Sigma #P8833) for 3 days, 5 mg/ml blasticidin S (Sigma #15205) for 5 days or 1 mg/ml G418 (Sigma #G8168) for 10 days until the complete death of mock-transfected cells.

    Techniques: Transfection, Selection, Staining, Plasmid Preparation

    Functionality of GFP gene from phRGFP plasmid is preserved in the products of recombination with pINS-Puro, pINS-Neo or pINS-Blast plasmids . HeLa cells transfected by 1 mkg of either Target vector phrGFP or the products of recombination (Blast × GFP, Neo × GFP and Puro × GFP). Cells transfected by the recombination products were selected by either blasticidin S, G418 or puromycin and stained by DAPI. Expression of the GFP was analyzed under the microscope. In case of transient transfection by phrGFP vector we usually observed 25% GFP positive cells. In contrast we observed that 80–100% of the cells transfected by the products of recombination and selected by the corresponding antibiotics are GFP positive. Scale bar: 20 μm.

    Journal: BMC Research Notes

    Article Title: A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination

    doi: 10.1186/1756-0500-1-135

    Figure Lengend Snippet: Functionality of GFP gene from phRGFP plasmid is preserved in the products of recombination with pINS-Puro, pINS-Neo or pINS-Blast plasmids . HeLa cells transfected by 1 mkg of either Target vector phrGFP or the products of recombination (Blast × GFP, Neo × GFP and Puro × GFP). Cells transfected by the recombination products were selected by either blasticidin S, G418 or puromycin and stained by DAPI. Expression of the GFP was analyzed under the microscope. In case of transient transfection by phrGFP vector we usually observed 25% GFP positive cells. In contrast we observed that 80–100% of the cells transfected by the products of recombination and selected by the corresponding antibiotics are GFP positive. Scale bar: 20 μm.

    Article Snippet: 24 hours after transfection media was replaced and the cells were either grown for another 24 hours (transient transfection) or selected with either 3 mg/ml puromycin (Sigma #P8833) for 3 days, 5 mg/ml blasticidin S (Sigma #15205) for 5 days or 1 mg/ml G418 (Sigma #G8168) for 10 days until the complete death of mock-transfected cells.

    Techniques: Plasmid Preparation, Transfection, Staining, Expressing, Microscopy

    Transient and permanent expression of the transferred DNA. (a) Overview of the experimental design to detect transient expression or stable integration of the transferred DNA. After infection of human cell lines with B. henselae , the DNA transferred through the T4SS will get to the nucleus where genes will be expressed. At 3 days postinfection, transient expression of gfp can be detected by flow cytometry. Antibiotic treatment was applied for long-term selection of neomycin-resistant colonies, to detect stable integration events. (b to d) Graphic representation of the percentage of GFP-positive cells obtained 3 days postinfection (b) and the number of G418-resistant colonies normalized for the number of cells at the beginning of the selection (c), as well as the Neo r /GFP + ratio (d). The different bars represented for each cell line correspond to the different relaxases under study, following the color code indicated in the squares at the top right. Data represent the means of the results from at least 3 independent experiments. Error bars indicate standard deviations. *, P

    Journal: Applied and Environmental Microbiology

    Article Title: The Conjugative Relaxase TrwC Promotes Integration of Foreign DNA in the Human Genome

    doi: 10.1128/AEM.00207-17

    Figure Lengend Snippet: Transient and permanent expression of the transferred DNA. (a) Overview of the experimental design to detect transient expression or stable integration of the transferred DNA. After infection of human cell lines with B. henselae , the DNA transferred through the T4SS will get to the nucleus where genes will be expressed. At 3 days postinfection, transient expression of gfp can be detected by flow cytometry. Antibiotic treatment was applied for long-term selection of neomycin-resistant colonies, to detect stable integration events. (b to d) Graphic representation of the percentage of GFP-positive cells obtained 3 days postinfection (b) and the number of G418-resistant colonies normalized for the number of cells at the beginning of the selection (c), as well as the Neo r /GFP + ratio (d). The different bars represented for each cell line correspond to the different relaxases under study, following the color code indicated in the squares at the top right. Data represent the means of the results from at least 3 independent experiments. Error bars indicate standard deviations. *, P

    Article Snippet: At 72 hpi, G418 disulfate salt (Sigma-Aldrich) was added to infected cells, and selection was maintained for 4 to 5 weeks.

    Techniques: Expressing, Infection, Flow Cytometry, Cytometry, Selection

    Sex selection and fitness of the flies carrying two copy of sex-sorter cassette. The sex-sorter cassette includes NeoR traF and PuroR dsxM genes that confer resistance to puromycin and geneticin antibiotics, respectively, in the sex-specific manner ( Figure 1a ). The PuroR dsxM gene is properly spliced and results in expression of the functional PuroR protein only in males, while NeoR traF expresses the NeoR protein only in females. To estimate the lowest concentration of an antibiotic at which male or female selection is enforced at 100%, the homozygous sex-sorter flies were raised on various concentrations of puromycin or geneticin. (a) Only male flies emerged from the food supplemented with 0.4 mg/mL or more of puromycin. (b) Raising the same flies on the food containing 0.2 mg/mL or more of geneticin resulted in the emergence of only female flies. To compare the fitness of homozygous sex-sorter flies to that of wild type ( wt ) flies, the embryo-to-adult survival of both fly types were compared under normal and selective conditions. (c) Embryos of both sex-sorter (gray bars) and wt flies (white bars) survived to the adulthood equally well on food without any antibiotics and died on the food supplemented with both puromycin and geneticin to 0.4 and 1.2 mg/mL. (d) The survival of male or female sex-sorter flies under selection treatments was statistically identical to that of the corresponding gender from wt flies raised under normal conditions. Bar plots show the average ± SD over at least three biological replicates. Statistical significance was estimated using a t test with equal variance. ( P ≥ 0.05 ns , P

    Journal: bioRxiv

    Article Title: A novel drug-inducible sex-separation technique for insects

    doi: 10.1101/2019.12.13.875716

    Figure Lengend Snippet: Sex selection and fitness of the flies carrying two copy of sex-sorter cassette. The sex-sorter cassette includes NeoR traF and PuroR dsxM genes that confer resistance to puromycin and geneticin antibiotics, respectively, in the sex-specific manner ( Figure 1a ). The PuroR dsxM gene is properly spliced and results in expression of the functional PuroR protein only in males, while NeoR traF expresses the NeoR protein only in females. To estimate the lowest concentration of an antibiotic at which male or female selection is enforced at 100%, the homozygous sex-sorter flies were raised on various concentrations of puromycin or geneticin. (a) Only male flies emerged from the food supplemented with 0.4 mg/mL or more of puromycin. (b) Raising the same flies on the food containing 0.2 mg/mL or more of geneticin resulted in the emergence of only female flies. To compare the fitness of homozygous sex-sorter flies to that of wild type ( wt ) flies, the embryo-to-adult survival of both fly types were compared under normal and selective conditions. (c) Embryos of both sex-sorter (gray bars) and wt flies (white bars) survived to the adulthood equally well on food without any antibiotics and died on the food supplemented with both puromycin and geneticin to 0.4 and 1.2 mg/mL. (d) The survival of male or female sex-sorter flies under selection treatments was statistically identical to that of the corresponding gender from wt flies raised under normal conditions. Bar plots show the average ± SD over at least three biological replicates. Statistical significance was estimated using a t test with equal variance. ( P ≥ 0.05 ns , P

    Article Snippet: A 1.1 g of the dry food was aliquoted per fly vial (FlyStaff.com) and mixed with 5 mL of distilled water supplemented with puromycin (Sigma #P8833) or geneticin (G418, Sigma #A1720), in varying concentrations from 0 to 1.2 mg/mL.

    Techniques: Selection, Expressing, Functional Assay, Concentration Assay

    Development of sex-sorter cassette in Drosophila . (a) Schematic maps of genetic constructs engineered and tested in the study. (b) Expression of antibiotic resistance genes ( PuroR and NeoR ) throughout Drosophila development confers resistance to puromycin and geneticin, respectively, supplemented on fly food. To insure that functional antibiotic-resistance proteins will be produced only in one or the other gender, sex-specific introns from two sex-determination genes ( tra and dsx ) were inserted into coding sequences of PuroR and NeoR . The entire sequences of female-specific traF and male-specific dsxM introns (highlighted in pink) are spliced out in the corresponding sex, but some sequences carrying a stop codon (TGA) are retained in the opposite sex ( Figure S2 ). The transgenic flies harboring one copy of a genetic construct were identified by a strong ubiquitous expression of dsRed (highlighted in purple). (c) Expression of Opie2-PuroR dsxM or Opie2-PuroR traF transgene rescues only transgenic males or females (red fluorescence) raised on the food supplemented with puromycin, while all wild type flies (no red fluorescence) and the transgenic flies of selected-out sex die during early development.

    Journal: bioRxiv

    Article Title: A novel drug-inducible sex-separation technique for insects

    doi: 10.1101/2019.12.13.875716

    Figure Lengend Snippet: Development of sex-sorter cassette in Drosophila . (a) Schematic maps of genetic constructs engineered and tested in the study. (b) Expression of antibiotic resistance genes ( PuroR and NeoR ) throughout Drosophila development confers resistance to puromycin and geneticin, respectively, supplemented on fly food. To insure that functional antibiotic-resistance proteins will be produced only in one or the other gender, sex-specific introns from two sex-determination genes ( tra and dsx ) were inserted into coding sequences of PuroR and NeoR . The entire sequences of female-specific traF and male-specific dsxM introns (highlighted in pink) are spliced out in the corresponding sex, but some sequences carrying a stop codon (TGA) are retained in the opposite sex ( Figure S2 ). The transgenic flies harboring one copy of a genetic construct were identified by a strong ubiquitous expression of dsRed (highlighted in purple). (c) Expression of Opie2-PuroR dsxM or Opie2-PuroR traF transgene rescues only transgenic males or females (red fluorescence) raised on the food supplemented with puromycin, while all wild type flies (no red fluorescence) and the transgenic flies of selected-out sex die during early development.

    Article Snippet: A 1.1 g of the dry food was aliquoted per fly vial (FlyStaff.com) and mixed with 5 mL of distilled water supplemented with puromycin (Sigma #P8833) or geneticin (G418, Sigma #A1720), in varying concentrations from 0 to 1.2 mg/mL.

    Techniques: Construct, Expressing, Functional Assay, Produced, Transgenic Assay, Fluorescence