smad7 antisense oligonucleotide custom lna oligonucleotide targeting smad7  (Qiagen)

 
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    Name:
    Custom LNA Oligonucleotide
    Description:
    Custom LNA Oligonucleotides are ideal for studies involving short or very similar sequences The high affinity of an LNA enhanced oligonucleotide to its complementary sequence results in dramatically improved specificity and sensitivity when compared with traditional DNA or RNA oligos In many cases LNA enhanced oligonucleotides can be used to distinguish between sequences differing by only a single nucleotide a feature that can be critical for the success of many experiments
    Catalog Number:
    339412
    Price:
    None
    Category:
    qPCR Arrays and Assays
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    Structured Review

    Qiagen smad7 antisense oligonucleotide custom lna oligonucleotide targeting smad7
    Custom LNA Oligonucleotide
    Custom LNA Oligonucleotides are ideal for studies involving short or very similar sequences The high affinity of an LNA enhanced oligonucleotide to its complementary sequence results in dramatically improved specificity and sensitivity when compared with traditional DNA or RNA oligos In many cases LNA enhanced oligonucleotides can be used to distinguish between sequences differing by only a single nucleotide a feature that can be critical for the success of many experiments
    https://www.bioz.com/result/smad7 antisense oligonucleotide custom lna oligonucleotide targeting smad7/product/Qiagen
    Average 96 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    smad7 antisense oligonucleotide custom lna oligonucleotide targeting smad7 - by Bioz Stars, 2020-07
    96/100 stars

    Images

    1) Product Images from "Smad7 knockdown activates protein kinase RNA-associated eIF2α pathway leading to colon cancer cell death"

    Article Title: Smad7 knockdown activates protein kinase RNA-associated eIF2α pathway leading to colon cancer cell death

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2017.103

    SMAD7 antisense (AS)-mediated DLD-1 cell death is reverted by PKR silencing. ( a ) Representative dot plots showing the percentages of AV- and/or PI-positive DLD-1 cells. Cells were transfected with either Smad7 sense (S) or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were incubated with either PKR-siRNA or scrambled-siRNA (both used at 100 nM) for further 24 h, and then washed with PBS and cultured for additional 48 h. ( b ) Representative histograms showing the percentage of cell death, as assessed by flow cytometry analysis of AV- and/or PI-positive cells, in DLD-1 cells treated as indicated in ( a ). Data are expressed as mean±S.E.M. of four experiments. P -values not significant are omitted
    Figure Legend Snippet: SMAD7 antisense (AS)-mediated DLD-1 cell death is reverted by PKR silencing. ( a ) Representative dot plots showing the percentages of AV- and/or PI-positive DLD-1 cells. Cells were transfected with either Smad7 sense (S) or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were incubated with either PKR-siRNA or scrambled-siRNA (both used at 100 nM) for further 24 h, and then washed with PBS and cultured for additional 48 h. ( b ) Representative histograms showing the percentage of cell death, as assessed by flow cytometry analysis of AV- and/or PI-positive cells, in DLD-1 cells treated as indicated in ( a ). Data are expressed as mean±S.E.M. of four experiments. P -values not significant are omitted

    Techniques Used: Transfection, Incubation, Cell Culture, Flow Cytometry, Cytometry

    SMAD7 antisense (AS)-induced PKR phosphorylation in DLD-1 cells does not rely on the modulation of known PKR-activating pathways. ( a–c ) DLD-1 cells were transfected with either Smad7 sense (S) or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, PKR ( a ), caspase-9, procaspase-3 and cleaved caspase-3 ( b ) as well as PACT expression ( c ) was assessed by western blotting. Staurosporine (ST) (1 μ g/ml) was used as a positive control for caspase-3 activation. β -Actin was used as loading control. One of three representative experiments is shown. ( d and e ) DLD-1 cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, RNA transcripts for the ER stress-related genes GRP-78 ( d ) and ATF6α ( e ) were determined by quantitative PCR. TM (1 μ g/ml) was used as a positive control. Levels are normalized to β -actin. Values mean±S.E.M. of three independent experiments. ( f ) DLD-1 cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, Smad7, p-PKR (Thr-446), GRP-78, ATF6 α and p-IRE1 α expression was evaluated by western blotting. β -Actin was used as a loading control. ( g ) DLD-1 cells were transfected with either CAD-11 AS oligonucleotide (used at 400 nM) or FSTL1 AS oligonucleotide (used at 10 nM) along with the respective negative controls (scrambled). After 24 h, cells were washed with PBS and cultured for further 30 min. CDH-11, FSTL1, p-PKR (Thr-446) and PKR expression was assessed by western blotting. β -Actin was used as a loading control. One of three representative experiments is shown
    Figure Legend Snippet: SMAD7 antisense (AS)-induced PKR phosphorylation in DLD-1 cells does not rely on the modulation of known PKR-activating pathways. ( a–c ) DLD-1 cells were transfected with either Smad7 sense (S) or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, PKR ( a ), caspase-9, procaspase-3 and cleaved caspase-3 ( b ) as well as PACT expression ( c ) was assessed by western blotting. Staurosporine (ST) (1 μ g/ml) was used as a positive control for caspase-3 activation. β -Actin was used as loading control. One of three representative experiments is shown. ( d and e ) DLD-1 cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, RNA transcripts for the ER stress-related genes GRP-78 ( d ) and ATF6α ( e ) were determined by quantitative PCR. TM (1 μ g/ml) was used as a positive control. Levels are normalized to β -actin. Values mean±S.E.M. of three independent experiments. ( f ) DLD-1 cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, Smad7, p-PKR (Thr-446), GRP-78, ATF6 α and p-IRE1 α expression was evaluated by western blotting. β -Actin was used as a loading control. ( g ) DLD-1 cells were transfected with either CAD-11 AS oligonucleotide (used at 400 nM) or FSTL1 AS oligonucleotide (used at 10 nM) along with the respective negative controls (scrambled). After 24 h, cells were washed with PBS and cultured for further 30 min. CDH-11, FSTL1, p-PKR (Thr-446) and PKR expression was assessed by western blotting. β -Actin was used as a loading control. One of three representative experiments is shown

    Techniques Used: Transfection, Expressing, Western Blot, Positive Control, Activation Assay, Real-time Polymerase Chain Reaction, Cell Culture

    Smad7 knockdown prevents PKR-p-58 IPK interaction in DLD-1 cells and activates the PKR/CHOP axis in human CRC explants. ( a ) Total proteins extracted from DLD-1 cells were immunoprecipitated by an anti-human Smad7 or control isotype (ve−) antibody and then subjected to immunoblotting analysis using PKR, p58 IPK and Smad7 antibodies. ( b ) Total proteins extracted from DLD-1 cells transfected with either Smad7 sense (S) or antisense (AS) oligonucleotide (both used at 2 μ g/ml) were immunoprecipitated by an anti-human PKR or control isotype (ve−) antibody and then subjected to immunoblotting analysis using p58 IPK , Smad7 and PKR antibodies. One of three representative experiments is shown. ( c ) Freshly obtained CRC explants were cultured in the presence of Smad7 S or AS oligonucleotide (both used at 8 μ g/ml) for 24 h. Smad7, p-PKR, PKR, CHOP, procaspase-3 and cleaved caspase-3 protein expression were assessed by western blotting. One of four representative experiments is shown. Right panels: Quantitative analysis of Smad7/ β -actin, p-PKR/PKR, CHOP/ β -actin and cleaved caspase-3/procaspase-3 protein ratio in extracts of CRC explants measured by densitometry scanning of western blots. Values are expressed in a.u. (arbitrary units) and are the means±S.E.M. of four experiments
    Figure Legend Snippet: Smad7 knockdown prevents PKR-p-58 IPK interaction in DLD-1 cells and activates the PKR/CHOP axis in human CRC explants. ( a ) Total proteins extracted from DLD-1 cells were immunoprecipitated by an anti-human Smad7 or control isotype (ve−) antibody and then subjected to immunoblotting analysis using PKR, p58 IPK and Smad7 antibodies. ( b ) Total proteins extracted from DLD-1 cells transfected with either Smad7 sense (S) or antisense (AS) oligonucleotide (both used at 2 μ g/ml) were immunoprecipitated by an anti-human PKR or control isotype (ve−) antibody and then subjected to immunoblotting analysis using p58 IPK , Smad7 and PKR antibodies. One of three representative experiments is shown. ( c ) Freshly obtained CRC explants were cultured in the presence of Smad7 S or AS oligonucleotide (both used at 8 μ g/ml) for 24 h. Smad7, p-PKR, PKR, CHOP, procaspase-3 and cleaved caspase-3 protein expression were assessed by western blotting. One of four representative experiments is shown. Right panels: Quantitative analysis of Smad7/ β -actin, p-PKR/PKR, CHOP/ β -actin and cleaved caspase-3/procaspase-3 protein ratio in extracts of CRC explants measured by densitometry scanning of western blots. Values are expressed in a.u. (arbitrary units) and are the means±S.E.M. of four experiments

    Techniques Used: Immunoprecipitation, Transfection, Cell Culture, Expressing, Western Blot

    Smad7 colocalizes and interacts with eIF2 α in colon cancer cells and controls eIF2 α downstream signaling. ( a ) Representative confocal laser scanning microscopy images showing Smad7 and eIF2 α colocalization in DLD-1 cell line. Scale bars, 25 μ m; scale bar inset, 10 μ m. ( b ) Total proteins extracted from DLD-1 cells were immunoprecipitated by an anti-human Smad7 or control isotype (ve−) antibody and then subjected to immunoblotting analysis using eIF2 α and Smad7 antibodies. One of three representative experiments in which similar results were obtained is shown. ( c ) Representative immunofluorescence pictures of DLD-1 cells showing that Smad7 knockdown enhances eIF2 α (Ser-51) phosphorylation. Cells were transfected with either Smad7 sense (S) or antisense (AS) oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were cultured for further 6 h, fixed and stained as described in ( a ). One of three representative experiments is shown. Scale bars, 20 μ m. ( d and e ) Representative immunofluorescence images of DLD-1 cells showing an increase of ATF4 and CHOP expression following Smad7 knockdown. Cells were transfected with either Smad7 S or AS (both used at 2 μ g/ml). After 24 h, cells were cultured for further 12 h, fixed and stained with DAPI nuclear staining (blue), anti-ATF4 or anti-CHOP and secondary Alexa Fluor 546 antibody (red). One of three representative experiments is shown. Scale bars, 20 μ m
    Figure Legend Snippet: Smad7 colocalizes and interacts with eIF2 α in colon cancer cells and controls eIF2 α downstream signaling. ( a ) Representative confocal laser scanning microscopy images showing Smad7 and eIF2 α colocalization in DLD-1 cell line. Scale bars, 25 μ m; scale bar inset, 10 μ m. ( b ) Total proteins extracted from DLD-1 cells were immunoprecipitated by an anti-human Smad7 or control isotype (ve−) antibody and then subjected to immunoblotting analysis using eIF2 α and Smad7 antibodies. One of three representative experiments in which similar results were obtained is shown. ( c ) Representative immunofluorescence pictures of DLD-1 cells showing that Smad7 knockdown enhances eIF2 α (Ser-51) phosphorylation. Cells were transfected with either Smad7 sense (S) or antisense (AS) oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were cultured for further 6 h, fixed and stained as described in ( a ). One of three representative experiments is shown. Scale bars, 20 μ m. ( d and e ) Representative immunofluorescence images of DLD-1 cells showing an increase of ATF4 and CHOP expression following Smad7 knockdown. Cells were transfected with either Smad7 S or AS (both used at 2 μ g/ml). After 24 h, cells were cultured for further 12 h, fixed and stained with DAPI nuclear staining (blue), anti-ATF4 or anti-CHOP and secondary Alexa Fluor 546 antibody (red). One of three representative experiments is shown. Scale bars, 20 μ m

    Techniques Used: Confocal Laser Scanning Microscopy, Immunoprecipitation, Immunofluorescence, Transfection, Cell Culture, Staining, Expressing

    Smad7 antisense (AS)-induced eIF2 α phosphorylation in DLD-1 cells relies on PKR activation. ( a ) DLD-1 cells were transfected with either Smad7 sense (S) or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were washed with PBS and cultured for 5, 15 and 30 min. p-PERK (Thr-981), p-GCN2 (Thr-899) and p-PKR (Thr-446) expression was assessed by western blotting. One of three representative experiments is shown. ( b ) Representative western blots for PKR in extracts of DLD-1 cells transfected with either scrambled-siRNA (100 nM) or increasing doses (50–100 nM) of PKR-siRNA for 24 h. ERK1 was used as a loading control. One of three representative experiments is shown. ( c ) Left panel: Cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were incubated with either PKR-siRNA or scrambled-siRNA (both used at 100 nM) for further 24 h and then washed with PBS and cultured for additional 6 h. Smad7, PKR, p-eIF2 α (Ser-51) and eIF2 α were assessed in extracts of DLD-1 cells by western blotting. β -Actin was used as a loading control. Right panel: Quantitative analysis of p-eIF2 α (Ser-51)/eIF2 α protein ratio in total extracts of DLD-1 cells as measured by densitometry scanning of western blots. Values are expressed in arbitrary units (a.u.) and indicate the mean±S.E.M. of three experiments. ( d and e ) Representative histograms showing the percentage of DLD1 cells expressing ATF4 ( d ) and CHOP ( e ). Cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were incubated with either PKR-siRNA or scrambled-siRNA (both used at 100 nM) for further 24 h and then washed with PBS and cultured for additional 12 h. Data are presented as mean values of positive cells per high power field (h.p.f.)±S.E.M. of three independent experiments, in which at least two sections per group were analyzed
    Figure Legend Snippet: Smad7 antisense (AS)-induced eIF2 α phosphorylation in DLD-1 cells relies on PKR activation. ( a ) DLD-1 cells were transfected with either Smad7 sense (S) or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were washed with PBS and cultured for 5, 15 and 30 min. p-PERK (Thr-981), p-GCN2 (Thr-899) and p-PKR (Thr-446) expression was assessed by western blotting. One of three representative experiments is shown. ( b ) Representative western blots for PKR in extracts of DLD-1 cells transfected with either scrambled-siRNA (100 nM) or increasing doses (50–100 nM) of PKR-siRNA for 24 h. ERK1 was used as a loading control. One of three representative experiments is shown. ( c ) Left panel: Cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were incubated with either PKR-siRNA or scrambled-siRNA (both used at 100 nM) for further 24 h and then washed with PBS and cultured for additional 6 h. Smad7, PKR, p-eIF2 α (Ser-51) and eIF2 α were assessed in extracts of DLD-1 cells by western blotting. β -Actin was used as a loading control. Right panel: Quantitative analysis of p-eIF2 α (Ser-51)/eIF2 α protein ratio in total extracts of DLD-1 cells as measured by densitometry scanning of western blots. Values are expressed in arbitrary units (a.u.) and indicate the mean±S.E.M. of three experiments. ( d and e ) Representative histograms showing the percentage of DLD1 cells expressing ATF4 ( d ) and CHOP ( e ). Cells were transfected with either Smad7 S or AS oligonucleotide (both used at 2 μ g/ml). After 24 h, cells were incubated with either PKR-siRNA or scrambled-siRNA (both used at 100 nM) for further 24 h and then washed with PBS and cultured for additional 12 h. Data are presented as mean values of positive cells per high power field (h.p.f.)±S.E.M. of three independent experiments, in which at least two sections per group were analyzed

    Techniques Used: Activation Assay, Transfection, Cell Culture, Expressing, Western Blot, Incubation

    Related Articles

    Transfection:

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    Article Title: Small RNA sequencing reveals miR-642a-3p as a novel adipocyte-specific microRNA and miR-30 as a key regulator of human adipogenesis
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    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
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    Reporter Assay:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Multiple Displacement Amplification:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Isolation:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Negative Control:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Labeling:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Modification:

    Article Title: Hodgkin Lymphoma Cell Lines Are Characterized by a Specific miRNA Expression Profile 1Hodgkin Lymphoma Cell Lines Are Characterized by a Specific miRNA Expression Profile 1 2
    Article Snippet: .. Using an Amaxa nucleofector device (AAD-1001; Amaxa, Gaithersburg, MD), cell lines were transfected with 2 µg of the modified psiCHECK2 plasmid containing a target 3′ UTR with or without 5.7 µM anti-miR-155 LNA-modified oligonucleotides (Custom LNA Oligonucleotides; Exiqon, Vedbaek, Denmark) in 100 µl of nucleofector solution (Amaxa) in triplicate. ..

    Inhibition:

    Article Title: Smad7 knockdown activates protein kinase RNA-associated eIF2α pathway leading to colon cancer cell death
    Article Snippet: .. Inhibition of Smad7 by Smad7 antisense oligonucleotide Custom LNA oligonucleotide targeting Smad7 was provided by Exiqon (Vedbaek, Denmark). .. Smad7 sense or Smad7 antisense oligonucleotide was used at the final concentration of 2 μ g/ml.

    Article Title: Smad7 knockdown activates protein kinase RNA-associated eIF2α pathway leading to colon cancer cell death
    Article Snippet: .. Inhibition of CAD-11 and FSTL1 by LNA oligonucleotides Custom LNA oligonucleotides targeting CAD-11 and FSTL1 were provided by Exiqon. .. HCT-116 and DLD-1 cells were transfected for 24 h with either CAD-11 or FSTL1 antisense, used at the final concentration of 400 and 10 nM, respectively, using Opti-MEM medium and Lipofectamine 3000 reagent (both from Life Technologies) according to the manufacturer's instructions.

    TBARS Assay:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Lactate Assay:

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

    Plasmid Preparation:

    Article Title: Hodgkin Lymphoma Cell Lines Are Characterized by a Specific miRNA Expression Profile 1Hodgkin Lymphoma Cell Lines Are Characterized by a Specific miRNA Expression Profile 1 2
    Article Snippet: .. Using an Amaxa nucleofector device (AAD-1001; Amaxa, Gaithersburg, MD), cell lines were transfected with 2 µg of the modified psiCHECK2 plasmid containing a target 3′ UTR with or without 5.7 µM anti-miR-155 LNA-modified oligonucleotides (Custom LNA Oligonucleotides; Exiqon, Vedbaek, Denmark) in 100 µl of nucleofector solution (Amaxa) in triplicate. ..

    Article Title: Suppression of Induced microRNA-15b Prevents Rapid Loss of Cardiac Function in a Dicer Depleted Model of Cardiac Dysfunction
    Article Snippet: .. Claycomb., Louisiana State University, New Orleans, LA; antibiotic (penicillin, streptomycin), tetramethylrhodamine methyl ester (TMRM) (Invitrogen Corporation, Carlsbad, CA); culture dishes (Nunc, Denmark); mirVana™ miRNA isolation kit, SYBR green-I (Ambion, Austin, TX); Taqman assays and Taqman Universal master mix (Applied Biosystems, Foster City, CA); FlexmiR™ MicroRNA Labeling Kit (Luminex,Austin,TX); JC-1 dye (Molecular Probes, Eugene, OR); anti-Pim1 antibody, anti-Dicer, anti-ANT1(Abcam, Cambridge, MA), anti-ANF (LS Biosciences,Seattle, WA); DharmaFECT 1 transfection reagent, mmu-miRNA-15b mimic and mimic control (Dharmacon RNA Technologies, Lafayette, CO); lactate assay kit-K627-100 (Biovision Research Products, CA); secondary anti-Rabbit IgG (Amersham Pharmacia Biotech, Piscataway, NJ); MDA standards (TBARS assay kit, ZeptoMetrix Corp., NY); Anti-mmu-miR-15b Custom LNA™ Oligonucleotide (Exiqon Inc., MA); Pim1–3′UTR plasmid (Signosis, Sunnyvale, CA); dual-luciferase reporter assay system (Promega, Madison, WI) miRCURY LNA™ microRNA Inhibitor Negative Control A (Exiqon Inc., MA). .. Development of the transgenic mice Mice homozygous for Dicer -floxed (loxP) alleles and transgenic B6129-Tg (Myh6-cre/Esr1)1Jmk/J were crossed to generate double transgenic (Myh6-cre/Esr1-Dicerfl/fl ) mice.

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