oligo  (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:
    Oligo dT Primer 50 µM
    Description:
    These are the same Ambion primers that are currently included in the RETROscript Kit SKU AM1710 They are provided at a stock concentration of 50 µM and are functionally tested using the RETROscript Kit
    Catalog Number:
    am5730g
    Price:
    None
    Applications:
    PCR & Real-Time PCR|RT-PCR|Reverse Transcription|Two-Step RT-PCR
    Category:
    Oligos Primers Probes Nucleotides
    Buy from Supplier


    Structured Review

    Thermo Fisher oligo
    RT-PCR confirmation of antisense transcription. a) A graphic representation of um02794 transcription. The grey line represents the genomic sequence (middle), the blue arrow represents predicted gene structure, and the red (top) and green (bottom) arrows represent sense and anti-sense ESTs respectively. The range of the genome coordinates was included. b) Detecting antisense transcripts corresponding to um02794 via strand specific RT-PCR. In lanes 2 to 5 first strand synthesis was carried out on <t>RNA</t> of CM grown haploid cells. In lanes 6 to 9 first strand synthesis was carried out on RNA of MN grown haploid cells. First synthesis reactions of lanes 2 and 6 were prepared using sense strand specific primers; lanes 3 and 7 anti sense specific primers; lane 4 and 8 <t>oligo</t> dT and lanes 5 and 9 DEPC-treated water. Lane 10 used genomic DNA from U. maydis strain 521 and lane 11 used water a PCR template. Lane 1 and 12: Full Ranger DNA ladder.
    These are the same Ambion primers that are currently included in the RETROscript Kit SKU AM1710 They are provided at a stock concentration of 50 µM and are functionally tested using the RETROscript Kit
    https://www.bioz.com/result/oligo/product/Thermo Fisher
    Average 99 stars, based on 1662 article reviews
    Price from $9.99 to $1999.99
    oligo - by Bioz Stars, 2020-11
    99/100 stars

    Images

    1) Product Images from "Gene discovery and transcript analyses in the corn smut pathogen Ustilago maydis: expressed sequence tag and genome sequence comparison"

    Article Title: Gene discovery and transcript analyses in the corn smut pathogen Ustilago maydis: expressed sequence tag and genome sequence comparison

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-8-334

    RT-PCR confirmation of antisense transcription. a) A graphic representation of um02794 transcription. The grey line represents the genomic sequence (middle), the blue arrow represents predicted gene structure, and the red (top) and green (bottom) arrows represent sense and anti-sense ESTs respectively. The range of the genome coordinates was included. b) Detecting antisense transcripts corresponding to um02794 via strand specific RT-PCR. In lanes 2 to 5 first strand synthesis was carried out on RNA of CM grown haploid cells. In lanes 6 to 9 first strand synthesis was carried out on RNA of MN grown haploid cells. First synthesis reactions of lanes 2 and 6 were prepared using sense strand specific primers; lanes 3 and 7 anti sense specific primers; lane 4 and 8 oligo dT and lanes 5 and 9 DEPC-treated water. Lane 10 used genomic DNA from U. maydis strain 521 and lane 11 used water a PCR template. Lane 1 and 12: Full Ranger DNA ladder.
    Figure Legend Snippet: RT-PCR confirmation of antisense transcription. a) A graphic representation of um02794 transcription. The grey line represents the genomic sequence (middle), the blue arrow represents predicted gene structure, and the red (top) and green (bottom) arrows represent sense and anti-sense ESTs respectively. The range of the genome coordinates was included. b) Detecting antisense transcripts corresponding to um02794 via strand specific RT-PCR. In lanes 2 to 5 first strand synthesis was carried out on RNA of CM grown haploid cells. In lanes 6 to 9 first strand synthesis was carried out on RNA of MN grown haploid cells. First synthesis reactions of lanes 2 and 6 were prepared using sense strand specific primers; lanes 3 and 7 anti sense specific primers; lane 4 and 8 oligo dT and lanes 5 and 9 DEPC-treated water. Lane 10 used genomic DNA from U. maydis strain 521 and lane 11 used water a PCR template. Lane 1 and 12: Full Ranger DNA ladder.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Sequencing, Polymerase Chain Reaction

    2) Product Images from "Apparent Defect in Yeast Bud-Site Selection Due to a Specific Failure to Splice the Pre-mRNA of a Regulator of Cell-Type-Specific Transcription"

    Article Title: Apparent Defect in Yeast Bud-Site Selection Due to a Specific Failure to Splice the Pre-mRNA of a Regulator of Cell-Type-Specific Transcription

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0047621

    Inefficient splicing of MATa1 pre-mRNA, but not of other pre-mRNAs, in bud13Δ and ist3Δ mutants. (A) Schematic representations of the genes tested and the primers used. Exons and introns are shown as open boxes and lines, respectively. Open arrowheads, forward and reverse primers corresponding to exon sequences; closed arrowheads, alternative forward primers corresponding to ACT1 and RPS17A intron sequences (see Table S1 ). (B–D) Analyses of spliced and unspliced mRNAs using the exon-derived primers. Cells of diploid strains YEF473 (wild-type), STY254 ( bud13Δ/bud13Δ ), STY260 ( ist3Δ/ist3Δ ), and STY464 ( pml1Δ/pml1Δ ) were grown in YM-P medium at 30°C to OD 600 ≈0.5. Total RNA was prepared, treated with DNase, and reverse-transcribed into single-stranded cDNA using oligo (dT) 16 primers (see Materials and Methods ). cDNAs were then amplified by PCR using the appropriate primers. RNA samples that were not treated with DNase (DNase-) and/or not subjected to reverse transcription (RT−), as indicated, were used as controls. Molecular-size markers were run in the outside lanes in each gel; their sizes are indicated. (E) Analysis of unspliced ACT1 and RPS17A transcripts using the intron-derived forward primers. The reverse primers and other conditions were as described for B–D. The arrow indicates the expected size for the cDNA derived from unspliced pre-mRNA (approximately the same for each gene). The other bands in the ACT1 lanes appear to be nonspecific PCR products.
    Figure Legend Snippet: Inefficient splicing of MATa1 pre-mRNA, but not of other pre-mRNAs, in bud13Δ and ist3Δ mutants. (A) Schematic representations of the genes tested and the primers used. Exons and introns are shown as open boxes and lines, respectively. Open arrowheads, forward and reverse primers corresponding to exon sequences; closed arrowheads, alternative forward primers corresponding to ACT1 and RPS17A intron sequences (see Table S1 ). (B–D) Analyses of spliced and unspliced mRNAs using the exon-derived primers. Cells of diploid strains YEF473 (wild-type), STY254 ( bud13Δ/bud13Δ ), STY260 ( ist3Δ/ist3Δ ), and STY464 ( pml1Δ/pml1Δ ) were grown in YM-P medium at 30°C to OD 600 ≈0.5. Total RNA was prepared, treated with DNase, and reverse-transcribed into single-stranded cDNA using oligo (dT) 16 primers (see Materials and Methods ). cDNAs were then amplified by PCR using the appropriate primers. RNA samples that were not treated with DNase (DNase-) and/or not subjected to reverse transcription (RT−), as indicated, were used as controls. Molecular-size markers were run in the outside lanes in each gel; their sizes are indicated. (E) Analysis of unspliced ACT1 and RPS17A transcripts using the intron-derived forward primers. The reverse primers and other conditions were as described for B–D. The arrow indicates the expected size for the cDNA derived from unspliced pre-mRNA (approximately the same for each gene). The other bands in the ACT1 lanes appear to be nonspecific PCR products.

    Techniques Used: Derivative Assay, Amplification, Polymerase Chain Reaction

    3) Product Images from "Progressive Tumor Features Accompany Epithelial-Mesenchymal Transition Induced in Mitochondrial DNA Depleted Cells"

    Article Title: Progressive Tumor Features Accompany Epithelial-Mesenchymal Transition Induced in Mitochondrial DNA Depleted Cells

    Journal: Cancer science

    doi: 10.1111/j.1349-7006.2008.00879.x

    Signal Transduction by TGFβ. A , RT-PCR for Type I TGFß Receptor. Total RNA was isolated from the indicated cells and one microgram was reverse transcribed using oligo dT. Primers were designed to amplify from 234 to 647 of Type I TGFβ
    Figure Legend Snippet: Signal Transduction by TGFβ. A , RT-PCR for Type I TGFß Receptor. Total RNA was isolated from the indicated cells and one microgram was reverse transcribed using oligo dT. Primers were designed to amplify from 234 to 647 of Type I TGFβ

    Techniques Used: Transduction, Reverse Transcription Polymerase Chain Reaction, Isolation

    4) Product Images from "Improved microarray gene expression profiling of virus-infected cells after removal of viral RNA"

    Article Title: Improved microarray gene expression profiling of virus-infected cells after removal of viral RNA

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-9-221

    vRNA depletion improves microarray analysis . Total RNA samples were obtained from mock- and MHV-infected LR7 cells at 6 h p.i. were subjected to the vRNA depletion protocol as detailed in the Materials and methods section. (A) Amplification of the mRNA was monitored by analyzing the cRNA samples with a Bioanalyzer. A representative mRNA amplification plot of a total RNA sample derived from MHV-infected cells at 6 h p.i. after vRNA removal is shown. The arrow indicates the marker peak. (B) Total RNA samples were treated with the biotinylated oligo's (indicated as vRNA depleted) and were processed for microarray analysis as described in the Methods section. The scatter plots display the average expression values from independent dye-swap hybridizations (n = 6) for each gene present on the arrays as described in legend of Fig.2. (C) The Venn diagrams show a comparison between the experiments with and without vRNA depletion.
    Figure Legend Snippet: vRNA depletion improves microarray analysis . Total RNA samples were obtained from mock- and MHV-infected LR7 cells at 6 h p.i. were subjected to the vRNA depletion protocol as detailed in the Materials and methods section. (A) Amplification of the mRNA was monitored by analyzing the cRNA samples with a Bioanalyzer. A representative mRNA amplification plot of a total RNA sample derived from MHV-infected cells at 6 h p.i. after vRNA removal is shown. The arrow indicates the marker peak. (B) Total RNA samples were treated with the biotinylated oligo's (indicated as vRNA depleted) and were processed for microarray analysis as described in the Methods section. The scatter plots display the average expression values from independent dye-swap hybridizations (n = 6) for each gene present on the arrays as described in legend of Fig.2. (C) The Venn diagrams show a comparison between the experiments with and without vRNA depletion.

    Techniques Used: Microarray, Infection, Amplification, Derivative Assay, Marker, Expressing

    5) Product Images from "Integrator mediates the biogenesis of enhancer RNAs"

    Article Title: Integrator mediates the biogenesis of enhancer RNAs

    Journal: Nature

    doi: 10.1038/nature14906

    EGF-induced eRNAs are predominantly non-polyadenylated a, We examined transcription at three enhancers adjacent to EGF-responsive genes CCNL1, NR4A1 and DUSP1. Total RNA samples were collected before and after EGF induction. Reverse transcription was performed with random hexamer primer or oligo d(T) primer. Each eRNA strand was analyzed by Real time PCR with specific primers. Error bars represent ± standard error of the mean (SEM, n=3 biological independent experiments). P
    Figure Legend Snippet: EGF-induced eRNAs are predominantly non-polyadenylated a, We examined transcription at three enhancers adjacent to EGF-responsive genes CCNL1, NR4A1 and DUSP1. Total RNA samples were collected before and after EGF induction. Reverse transcription was performed with random hexamer primer or oligo d(T) primer. Each eRNA strand was analyzed by Real time PCR with specific primers. Error bars represent ± standard error of the mean (SEM, n=3 biological independent experiments). P

    Techniques Used: Random Hexamer Labeling, Real-time Polymerase Chain Reaction

    6) Product Images from "Sensitivity of 70-mer oligonucleotides and cDNAs for microarray analysis of gene expression in Arabidopsis and its related species"

    Article Title: Sensitivity of 70-mer oligonucleotides and cDNAs for microarray analysis of gene expression in Arabidopsis and its related species

    Journal: Plant biotechnology journal

    doi: 10.1046/j.1467-7652.2003.00048.x

    Bar plots comparing gene expression changes between Arabidopsis thaliana leaves and flower buds detected by microarrays with ‘undenatured’ cDNA features, undenatured oligo features, and quantitative RT-PCR (QRT-PCR). Results show fold-change of expression ratios (flower buds vs. leaves; F/L) obtained from six replications in the cDNA and oligo microarray and 3 replications from QRT-PCR, and are grouped into four categories: (a) similar results for cDNA, oligo and QRT-PCR (b) similar for oligos and QRT-PCR, but different for cDNA (c) different for cDNA and oligos, but similar for cDNA and QRT-PCR (d) similar result for cDNA and oligo but different for QRT-PCR.
    Figure Legend Snippet: Bar plots comparing gene expression changes between Arabidopsis thaliana leaves and flower buds detected by microarrays with ‘undenatured’ cDNA features, undenatured oligo features, and quantitative RT-PCR (QRT-PCR). Results show fold-change of expression ratios (flower buds vs. leaves; F/L) obtained from six replications in the cDNA and oligo microarray and 3 replications from QRT-PCR, and are grouped into four categories: (a) similar results for cDNA, oligo and QRT-PCR (b) similar for oligos and QRT-PCR, but different for cDNA (c) different for cDNA and oligos, but similar for cDNA and QRT-PCR (d) similar result for cDNA and oligo but different for QRT-PCR.

    Techniques Used: Expressing, Quantitative RT-PCR, Microarray

    Related Articles

    Co-Immunoprecipitation Assay:

    Article Title: Translation of yes-associated protein (YAP) was antagonized by its circular RNA via suppressing the assembly of the translation initiation machinery
    Article Snippet: .. In brief, the cells were lysed in co-IP buffer and then incubated with 3 μg biotinylated DNA oligo probes against circ-Yap or Yap mRNA at room temperature for 2 h. Fifty microliters of Streptavidin C1 magnetic beads (Invitrogen) were added to each binding reaction and further incubated at room temperature for another 1 h. The beads were washed briefly with co-IP buffer for five times. .. The bound proteins in the pull-down material were analyzed by western blotting.

    Synthesized:

    Article Title: Down-regulation of TERE1/UBIAD1 activated Ras-MAPK signalling and induced cell proliferation
    Article Snippet: .. SiRNA oligos were synthesized by Invitrogen as follows: Negative control siRNA, 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) 5′-ACGUGACACGUUCGGAGAATT-3′(antisense); TERE1-291 siRNA, 5′-GUAAUUUGGUCAACACUUATT-3′(sense) 5′-UAAGUGUUGACCAAAUUACCG3-′(antisense); TERE1-22 siRNA, 5′-CAAGGGCAUUGACCACAAATT-3′ (sense) 5′-UUUGUGGUCAAUGCCCUUGGA-3′ (antisense) For each transfection, oligomer–Lipofectamine™ 2000 complexes were prepared by mixing siRNA oligos with Lipofectamine™ 2000 and added to each well containing cells and medium. ..

    Magnetic Beads:

    Article Title: Translation of yes-associated protein (YAP) was antagonized by its circular RNA via suppressing the assembly of the translation initiation machinery
    Article Snippet: .. In brief, the cells were lysed in co-IP buffer and then incubated with 3 μg biotinylated DNA oligo probes against circ-Yap or Yap mRNA at room temperature for 2 h. Fifty microliters of Streptavidin C1 magnetic beads (Invitrogen) were added to each binding reaction and further incubated at room temperature for another 1 h. The beads were washed briefly with co-IP buffer for five times. .. The bound proteins in the pull-down material were analyzed by western blotting.

    Transfection:

    Article Title: Concordant and Discordant Regulation of Target Genes by miR-31 and Its Isoforms
    Article Snippet: .. The level of overexpressed isomiR-31s in MCF-7 cells transfected with synthetic oligos from Ambion (B) and Dharmacon (C). .. The expression level was shown as miR-31 (−ΔCt), which is equal to – (CtmiR-31 −CtU6). (TIF).

    Article Title: Down-regulation of TERE1/UBIAD1 activated Ras-MAPK signalling and induced cell proliferation
    Article Snippet: .. SiRNA oligos were synthesized by Invitrogen as follows: Negative control siRNA, 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) 5′-ACGUGACACGUUCGGAGAATT-3′(antisense); TERE1-291 siRNA, 5′-GUAAUUUGGUCAACACUUATT-3′(sense) 5′-UAAGUGUUGACCAAAUUACCG3-′(antisense); TERE1-22 siRNA, 5′-CAAGGGCAUUGACCACAAATT-3′ (sense) 5′-UUUGUGGUCAAUGCCCUUGGA-3′ (antisense) For each transfection, oligomer–Lipofectamine™ 2000 complexes were prepared by mixing siRNA oligos with Lipofectamine™ 2000 and added to each well containing cells and medium. ..

    Ligation:

    Article Title: Structural bias in T4 RNA ligase-mediated 3?-adapter ligation
    Article Snippet: .. To assess the frequency of nucleotides at each randomized position in the oligo pool, the random RNA oligos were subjected to Ion Torrent sequencing without undergoing adapter ligation ( ). ..

    Incubation:

    Article Title: HuR stabilizes HTT mRNA via interacting with its exon 11 in a mutant HTT-dependent manner
    Article Snippet: .. Then the titrated HuR protein (2 mg/mL) and RNA oligos were dissolved in the EMSA interaction buffer (1 × R-EMSA Binding Buffer (Thermo Fisher Scientific, #20158) + 5% glycerol + 100 μg/mL tRNA+1× RNase inhibitor (Thermo Fisher Scientific, #N8080119)), and then incubated for 30 min at room temperature (25 °C). .. The reaction mixture was then loaded onto a 6% acrylamide native gel and run the electrophoresis at 4 °C.

    Article Title: Translation of yes-associated protein (YAP) was antagonized by its circular RNA via suppressing the assembly of the translation initiation machinery
    Article Snippet: .. In brief, the cells were lysed in co-IP buffer and then incubated with 3 μg biotinylated DNA oligo probes against circ-Yap or Yap mRNA at room temperature for 2 h. Fifty microliters of Streptavidin C1 magnetic beads (Invitrogen) were added to each binding reaction and further incubated at room temperature for another 1 h. The beads were washed briefly with co-IP buffer for five times. .. The bound proteins in the pull-down material were analyzed by western blotting.

    other:

    Article Title: Enrichment of small pathogenic deletions at chromosome 9p24.3 and 9q34.3 involving DOCK8, KANK1, EHMT1 genes identified by using high-resolution oligonucleotide-single nucleotide polymorphism array analysis
    Article Snippet: Oligonucleotide-single nucleotide polymorphism array analysis platforms and threshold setting Oligo-SNP array analysis was performed on either Human SNP Array 6.0 (in 2011) or CytoScan® HD (2012–2015)(Affymetrix, Santa Clara, CA), using genomic DNA extracted from whole blood.

    Negative Control:

    Article Title: Down-regulation of TERE1/UBIAD1 activated Ras-MAPK signalling and induced cell proliferation
    Article Snippet: .. SiRNA oligos were synthesized by Invitrogen as follows: Negative control siRNA, 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) 5′-ACGUGACACGUUCGGAGAATT-3′(antisense); TERE1-291 siRNA, 5′-GUAAUUUGGUCAACACUUATT-3′(sense) 5′-UAAGUGUUGACCAAAUUACCG3-′(antisense); TERE1-22 siRNA, 5′-CAAGGGCAUUGACCACAAATT-3′ (sense) 5′-UUUGUGGUCAAUGCCCUUGGA-3′ (antisense) For each transfection, oligomer–Lipofectamine™ 2000 complexes were prepared by mixing siRNA oligos with Lipofectamine™ 2000 and added to each well containing cells and medium. ..

    Expressing:

    Article Title: Hedgehog Signaling and Bmi-1 Regulate Self-renewal of Normal and Malignant Human Mammary Stem Cells
    Article Snippet: .. Three human Bmi-1 (hBmi-1) siRNA oligos were purchased from Ambion, Inc. (Austin, TX; Silencer Predesigned siRNAs) and were used to confirm the knockdown of Bmi-1 expression in primary human mammary epithelial cells. .. All the siRNA sequences were converted to small hairpins (shRNA) and inserted into lentivirus vector LentiLox 3.7.

    Sequencing:

    Article Title: Structural bias in T4 RNA ligase-mediated 3?-adapter ligation
    Article Snippet: .. To assess the frequency of nucleotides at each randomized position in the oligo pool, the random RNA oligos were subjected to Ion Torrent sequencing without undergoing adapter ligation ( ). ..

    Binding Assay:

    Article Title: HuR stabilizes HTT mRNA via interacting with its exon 11 in a mutant HTT-dependent manner
    Article Snippet: .. Then the titrated HuR protein (2 mg/mL) and RNA oligos were dissolved in the EMSA interaction buffer (1 × R-EMSA Binding Buffer (Thermo Fisher Scientific, #20158) + 5% glycerol + 100 μg/mL tRNA+1× RNase inhibitor (Thermo Fisher Scientific, #N8080119)), and then incubated for 30 min at room temperature (25 °C). .. The reaction mixture was then loaded onto a 6% acrylamide native gel and run the electrophoresis at 4 °C.

    Article Title: Translation of yes-associated protein (YAP) was antagonized by its circular RNA via suppressing the assembly of the translation initiation machinery
    Article Snippet: .. In brief, the cells were lysed in co-IP buffer and then incubated with 3 μg biotinylated DNA oligo probes against circ-Yap or Yap mRNA at room temperature for 2 h. Fifty microliters of Streptavidin C1 magnetic beads (Invitrogen) were added to each binding reaction and further incubated at room temperature for another 1 h. The beads were washed briefly with co-IP buffer for five times. .. The bound proteins in the pull-down material were analyzed by western blotting.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher sirna oligos
    GGTase3 geranylgeranylates FBXL2 and is required for its localization to cellular membranes. (a) Recombinant GGtase3 geranylgeranylates purified FBXL2. Indicated amounts of purified FBXL2 were incubated with 100 ng of purified GGTase3 (either tagged [T] or untagged [UT] versions) to carry out in vitro geranylgeranylation assay using saturating concentrations of tritiated [H 3 ]-GGPP as described in methods. Each data point represents mean+/− SD of three biological replicates. Michaelis-Menten kinetics was used to generate an apparent K m value of 1.2μM using Prism Graphpad software. (b) In vitro geranylgeranylation assay was carried out and measured as in (a) using 10 μM of purified FBXL2, FBXW7, or K-RAS4B and 100 ng of purified GGTase3. Bar graphs represent mean +/− SD from three biological replicates. Source data for panels a and b are available with the paper online. (c) RPE1-HTERT cells were cotransfected with the indicated plasmids and processed for the detection of geranylgeranylated FBXL2 using a “Click-IT” assay, as described in methods. The experiment was repeated three times. Representative result is shown. Uncropped blot/gel images are shown in Supplementary Data Set 1 . (d) HeLa cells were transfected with the indicated <t>siRNA</t> <t>oligos</t> and cDNAs. Twenty-four hours post-transfection cells were incubated with geranylgeranyl-azide for 16 hours, harvested, lysed, and azide selective ligation reaction with sDIBO-Biotin was performed for one hour to label geranylgeranylated proteins via copper-free “Click-IT” reaction. After immunoprecipitation with an anti-HA antibody, immunoblots were carried out. The experiment was repeated four times. Representative result is shown. Uncropped blot/gel images are shown in Supplementary Data Set 1 . (e) HeLa cells were transfected first with the indicated siRNA oligos and then with the indicated GFP-tagged proteins. Live cell confocal imaging was carried out as described in methods. Images show representative frames of three independent experiments. Bar size: 10 μm.
    Sirna Oligos, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna oligos/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    sirna oligos - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    97
    Thermo Fisher biotinylated probe
    HBM mutations do not impact DNA-binding by MYC:MAX complexes. ( A ) Recombinant MYC (WT, 4A, or VP16 HBM):MAX dimers used in these assays. Dimers were separated by SDS-PAGE alongside a BSA standard, and stained using Coomassie Brilliant Blue. ( B ) Electrophoretic mobility shift assay (EMSA) using recombinant MYC:MAX and MAX:MAX dimers incubated with a <t>biotinylated</t> E-box probe (5′-GCTCAGGGACCACGTGGTCGGGGATC-3′). Binding reactions were conducted with or without 100-fold excess of unlabeled specific (as above) or non-specific (5′-GCTCAGGGACCA GC TGGTCGGGGATC-3′) competitors.
    Biotinylated Probe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated probe/product/Thermo Fisher
    Average 97 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    biotinylated probe - by Bioz Stars, 2020-11
    97/100 stars
      Buy from Supplier

    Image Search Results


    GGTase3 geranylgeranylates FBXL2 and is required for its localization to cellular membranes. (a) Recombinant GGtase3 geranylgeranylates purified FBXL2. Indicated amounts of purified FBXL2 were incubated with 100 ng of purified GGTase3 (either tagged [T] or untagged [UT] versions) to carry out in vitro geranylgeranylation assay using saturating concentrations of tritiated [H 3 ]-GGPP as described in methods. Each data point represents mean+/− SD of three biological replicates. Michaelis-Menten kinetics was used to generate an apparent K m value of 1.2μM using Prism Graphpad software. (b) In vitro geranylgeranylation assay was carried out and measured as in (a) using 10 μM of purified FBXL2, FBXW7, or K-RAS4B and 100 ng of purified GGTase3. Bar graphs represent mean +/− SD from three biological replicates. Source data for panels a and b are available with the paper online. (c) RPE1-HTERT cells were cotransfected with the indicated plasmids and processed for the detection of geranylgeranylated FBXL2 using a “Click-IT” assay, as described in methods. The experiment was repeated three times. Representative result is shown. Uncropped blot/gel images are shown in Supplementary Data Set 1 . (d) HeLa cells were transfected with the indicated siRNA oligos and cDNAs. Twenty-four hours post-transfection cells were incubated with geranylgeranyl-azide for 16 hours, harvested, lysed, and azide selective ligation reaction with sDIBO-Biotin was performed for one hour to label geranylgeranylated proteins via copper-free “Click-IT” reaction. After immunoprecipitation with an anti-HA antibody, immunoblots were carried out. The experiment was repeated four times. Representative result is shown. Uncropped blot/gel images are shown in Supplementary Data Set 1 . (e) HeLa cells were transfected first with the indicated siRNA oligos and then with the indicated GFP-tagged proteins. Live cell confocal imaging was carried out as described in methods. Images show representative frames of three independent experiments. Bar size: 10 μm.

    Journal: Nature structural & molecular biology

    Article Title: GGTase3 is a Newly Identified Geranylgeranyltransferase Targeting a Ubiquitin Ligase

    doi: 10.1038/s41594-019-0249-3

    Figure Lengend Snippet: GGTase3 geranylgeranylates FBXL2 and is required for its localization to cellular membranes. (a) Recombinant GGtase3 geranylgeranylates purified FBXL2. Indicated amounts of purified FBXL2 were incubated with 100 ng of purified GGTase3 (either tagged [T] or untagged [UT] versions) to carry out in vitro geranylgeranylation assay using saturating concentrations of tritiated [H 3 ]-GGPP as described in methods. Each data point represents mean+/− SD of three biological replicates. Michaelis-Menten kinetics was used to generate an apparent K m value of 1.2μM using Prism Graphpad software. (b) In vitro geranylgeranylation assay was carried out and measured as in (a) using 10 μM of purified FBXL2, FBXW7, or K-RAS4B and 100 ng of purified GGTase3. Bar graphs represent mean +/− SD from three biological replicates. Source data for panels a and b are available with the paper online. (c) RPE1-HTERT cells were cotransfected with the indicated plasmids and processed for the detection of geranylgeranylated FBXL2 using a “Click-IT” assay, as described in methods. The experiment was repeated three times. Representative result is shown. Uncropped blot/gel images are shown in Supplementary Data Set 1 . (d) HeLa cells were transfected with the indicated siRNA oligos and cDNAs. Twenty-four hours post-transfection cells were incubated with geranylgeranyl-azide for 16 hours, harvested, lysed, and azide selective ligation reaction with sDIBO-Biotin was performed for one hour to label geranylgeranylated proteins via copper-free “Click-IT” reaction. After immunoprecipitation with an anti-HA antibody, immunoblots were carried out. The experiment was repeated four times. Representative result is shown. Uncropped blot/gel images are shown in Supplementary Data Set 1 . (e) HeLa cells were transfected first with the indicated siRNA oligos and then with the indicated GFP-tagged proteins. Live cell confocal imaging was carried out as described in methods. Images show representative frames of three independent experiments. Bar size: 10 μm.

    Article Snippet: Cells were either with siRNA oligos and/or plasmids for the times indicated in figure legends using Lipofactemine®3000 reagent for 1 hour at 37°C prior to metabolic labeling with geranylgeranly azide (30uM, Thermo Scientific # C10249) for 24 hours in DMEM medium supplemented with 10% dialyzed FBS and 5 mM sodium pyruvate.

    Techniques: Recombinant, Purification, Incubation, In Vitro, Software, Transfection, Ligation, Immunoprecipitation, Western Blot, Imaging

    Induction of SASP by activating the cytoplasmic DNA sensing pathway. a Senescent TIG-3 cells induced by oncogenic Ras expression (lane 2–4) were transfected with previously validated siRNA oligos indicated at the top of the panel for twice at 2 day intervals. These cells were then subjected to western blotting using antibodies shown right ( a ), RT-qPCR analysis of SASP factor gene expression ( b ), analysis of intracellular ROS levels ( c ) or immunofluorescence staining for markers of DNA damage (γ-H2AX [red], phosphor-Ser/Thr ATM/ATR (pST/Q) substrate [green] and 40,6-diamidino-2-phenylindole [blue]) on day 4 ( d ). The representative data from three independent experiments are shown. Tubulin was used as a loading control ( a ). For all graphs, error bars indicate mean ± standard deviation (s.d.) of triplicate measurements. (* P

    Journal: Nature Communications

    Article Title: Downregulation of cytoplasmic DNases is implicated in cytoplasmic DNA accumulation and SASP in senescent cells

    doi: 10.1038/s41467-018-03555-8

    Figure Lengend Snippet: Induction of SASP by activating the cytoplasmic DNA sensing pathway. a Senescent TIG-3 cells induced by oncogenic Ras expression (lane 2–4) were transfected with previously validated siRNA oligos indicated at the top of the panel for twice at 2 day intervals. These cells were then subjected to western blotting using antibodies shown right ( a ), RT-qPCR analysis of SASP factor gene expression ( b ), analysis of intracellular ROS levels ( c ) or immunofluorescence staining for markers of DNA damage (γ-H2AX [red], phosphor-Ser/Thr ATM/ATR (pST/Q) substrate [green] and 40,6-diamidino-2-phenylindole [blue]) on day 4 ( d ). The representative data from three independent experiments are shown. Tubulin was used as a loading control ( a ). For all graphs, error bars indicate mean ± standard deviation (s.d.) of triplicate measurements. (* P

    Article Snippet: RNAi was performed by the transfection of siRNA oligos using the Lipofectamine™ RNAiMAX transfection reagent (Thermo Fisher Scientific), according to the manufacturer’s instructions.

    Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Standard Deviation

    The knockdown of cytoplasmic DNases activates the IFN-β pathway. a – c Pre-senescent TIG-3 cells were subjected to transfection with indicated siRNA oligos twice (at 2 day intervals). These cells were then subjected to western blotting using antibodies shown right ( a ), isolation of cytoplasmic fraction followed by qPCR analysis of chromosomal DNA ( b ) or qPCR analysis of SASP factor gene expression ( c ). Tubulin was used as a loading control ( a ). The representative data from three independent experiments are shown. For all graphs, error bars indicate mean ± standard deviation (s.d.) of triplicate measurements. (** P

    Journal: Nature Communications

    Article Title: Downregulation of cytoplasmic DNases is implicated in cytoplasmic DNA accumulation and SASP in senescent cells

    doi: 10.1038/s41467-018-03555-8

    Figure Lengend Snippet: The knockdown of cytoplasmic DNases activates the IFN-β pathway. a – c Pre-senescent TIG-3 cells were subjected to transfection with indicated siRNA oligos twice (at 2 day intervals). These cells were then subjected to western blotting using antibodies shown right ( a ), isolation of cytoplasmic fraction followed by qPCR analysis of chromosomal DNA ( b ) or qPCR analysis of SASP factor gene expression ( c ). Tubulin was used as a loading control ( a ). The representative data from three independent experiments are shown. For all graphs, error bars indicate mean ± standard deviation (s.d.) of triplicate measurements. (** P

    Article Snippet: RNAi was performed by the transfection of siRNA oligos using the Lipofectamine™ RNAiMAX transfection reagent (Thermo Fisher Scientific), according to the manufacturer’s instructions.

    Techniques: Transfection, Western Blot, Isolation, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

    HBM mutations do not impact DNA-binding by MYC:MAX complexes. ( A ) Recombinant MYC (WT, 4A, or VP16 HBM):MAX dimers used in these assays. Dimers were separated by SDS-PAGE alongside a BSA standard, and stained using Coomassie Brilliant Blue. ( B ) Electrophoretic mobility shift assay (EMSA) using recombinant MYC:MAX and MAX:MAX dimers incubated with a biotinylated E-box probe (5′-GCTCAGGGACCACGTGGTCGGGGATC-3′). Binding reactions were conducted with or without 100-fold excess of unlabeled specific (as above) or non-specific (5′-GCTCAGGGACCA GC TGGTCGGGGATC-3′) competitors.

    Journal: bioRxiv

    Article Title: MYC regulates ribosome biogenesis and mitochondrial gene expression programs through interaction with Host Cell Factor-1

    doi: 10.1101/2020.06.22.164764

    Figure Lengend Snippet: HBM mutations do not impact DNA-binding by MYC:MAX complexes. ( A ) Recombinant MYC (WT, 4A, or VP16 HBM):MAX dimers used in these assays. Dimers were separated by SDS-PAGE alongside a BSA standard, and stained using Coomassie Brilliant Blue. ( B ) Electrophoretic mobility shift assay (EMSA) using recombinant MYC:MAX and MAX:MAX dimers incubated with a biotinylated E-box probe (5′-GCTCAGGGACCACGTGGTCGGGGATC-3′). Binding reactions were conducted with or without 100-fold excess of unlabeled specific (as above) or non-specific (5′-GCTCAGGGACCA GC TGGTCGGGGATC-3′) competitors.

    Article Snippet: For reactions involving unlabeled specific or non-specific competitor, these were included in the binding reaction at a 100-fold excess over the biotinylated probe.

    Techniques: Binding Assay, Recombinant, SDS Page, Staining, Electrophoretic Mobility Shift Assay, Incubation

    Enzymatic addition of a single 3′ biotin moiety to unmodified oligos Unmodified oligos were biotinylated with biotin-ddUTP and terminal deoxynucleotidyl transferase (TdT). Biotinylation efficiency was assessed by Urea PAGE. Successful biotinylation causes an upward shift of the oligo in the gel. In each lane, 1 pmol of oligo was loaded. Left: Biotinylation efficiency assessment for single-probe reactions, a representative gel is shown. Right: Biotinylation efficiency assessment for the pool of 15 oligos. L = DNA ladder.

    Journal: bioRxiv

    Article Title: Ribo-Pop: Simple, cost-effective, and widely applicable ribosomal RNA depletion

    doi: 10.1101/2020.05.19.102293

    Figure Lengend Snippet: Enzymatic addition of a single 3′ biotin moiety to unmodified oligos Unmodified oligos were biotinylated with biotin-ddUTP and terminal deoxynucleotidyl transferase (TdT). Biotinylation efficiency was assessed by Urea PAGE. Successful biotinylation causes an upward shift of the oligo in the gel. In each lane, 1 pmol of oligo was loaded. Left: Biotinylation efficiency assessment for single-probe reactions, a representative gel is shown. Right: Biotinylation efficiency assessment for the pool of 15 oligos. L = DNA ladder.

    Article Snippet: To minimize the effect of pipetting error on probe efficacy measurements, each replicate uses a different sample of larval RNA and an independent dilution of biotinylated probe.

    Techniques: Polyacrylamide Gel Electrophoresis