quant it rna assay kit  (Thermo Fisher)


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    Structured Review

    Thermo Fisher quant it rna assay kit
    Quant It Rna Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quant it rna assay kit/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    quant it rna assay kit - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Electrophoresis:

    Article Title: Deficit of Kcnma1 mRNA expression in the dentate gyrus of epileptic rats
    Article Snippet: The concentration and purity of total RNA for each sample was determined by the Quant-iT RNA Assay Kit and Q32857 Qubit fluorometer (Carlsbad, Invitrogen, California, USA) and confirmed by optical density measurements at 260 and 280nm using a BioMate 5 UV-visible spectrophotometer (Thermo Spectronic, Waltham, Massachusetts, USA). .. The integrity of the extracted RNA was confirmed by electrophoresis under denaturating condition [ ].

    Centrifugation:

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: During the process of RNA purification, genomic DNA was removed effectively in a single centrifugation step. .. RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm.

    Amplification:

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm. .. The RT reaction mixture was then diluted 1:10 with nuclease-free water and used for PCR amplification of MMP cDNA in the presence of the primers.

    Synthesized:

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm. .. First-strand cDNA (cDNA) was synthesized with the iScript™ cDNA synthesis kit (Bio-Rad, Hercules, CA, USA) using 20 μl reaction mixture containing 0.25 μg total RNA, 4 μl 5× iScript reaction mixture, and 1 μl iScript RT.

    Article Title: Identification of the Efflux Transporter of the Fluoroquinolone Antibiotic Ciprofloxacin in Murine Macrophages: Studies with Ciprofloxacin-Resistant Cells ▿
    Article Snippet: .. Total RNA was purified with Turbo DNase (Ambion, Austin, TX), and the concentration was measured with a Qubit fluorimeter, using a Quant-iT RNA assay kit (Invitrogen). cDNA was synthesized using 1 μg of total purified RNA and random hexamer primers (Promega reverse transcription system; Promega Co., Madison, WI). .. Primer pairs for all Mrp (also referred to as Abcc ) genes investigated were designed using Primer 3 and Autoprime programs , taking into account exon-intron boundaries to prevent genomic DNA amplification.

    Isolation:

    Article Title: Deficit of Kcnma1 mRNA expression in the dentate gyrus of epileptic rats
    Article Snippet: Tissue was collected, weighed (about 20 mg), homogenized, and processed for total RNA isolation at 4°C using the RNAqueous-4PCR Kit (Foster City, California, USA), following manufacturer's instructions. .. The concentration and purity of total RNA for each sample was determined by the Quant-iT RNA Assay Kit and Q32857 Qubit fluorometer (Carlsbad, Invitrogen, California, USA) and confirmed by optical density measurements at 260 and 280nm using a BioMate 5 UV-visible spectrophotometer (Thermo Spectronic, Waltham, Massachusetts, USA).

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: Total RNA was isolated from cells using the RNeasy Plus Mini kit (Qiagen, Santa Clarita, CA, USA). .. RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm.

    Article Title: Integrin-linked kinase regulates smooth muscle differentiation marker gene expression in airway tissue
    Article Snippet: Total RNA was isolated from the pulverized muscle extracts using the RNeasy Mini Kit (Qiagen, Chatsworth, CA) according to the manufacturer's instructions. .. The eluted RNA was quantified using the Quant-iT RNA Assay Kit (Invitrogen).

    Article Title: Identification and characterization of the doublesex gene of Nasonia
    Article Snippet: Poly(A) RNA was isolated using the Dynabeads mRNA DIRECT Kit (Dynal Biotech, Norway) by the mini volumes protocol. .. When necessary, RNA was quantified using a Qubit fluorometer (Invitrogen, CA) and a Quant-iT RNA Assay Kit (Invitrogen, CA) or a ND-1000 Spectrophotometer (Nanodrop technologies, UK).

    Article Title: Identification of the Efflux Transporter of the Fluoroquinolone Antibiotic Ciprofloxacin in Murine Macrophages: Studies with Ciprofloxacin-Resistant Cells ▿
    Article Snippet: Paragraph title: Isolation of RNA and synthesis of cDNA. ... Total RNA was purified with Turbo DNase (Ambion, Austin, TX), and the concentration was measured with a Qubit fluorimeter, using a Quant-iT RNA assay kit (Invitrogen). cDNA was synthesized using 1 μg of total purified RNA and random hexamer primers (Promega reverse transcription system; Promega Co., Madison, WI).

    Article Title: Semen miRNAs Contained in Exosomes as Non-Invasive Biomarkers for Prostate Cancer Diagnosis
    Article Snippet: Total RNA was obtained from exosomes using the miRCURY RNA Isolation Kit-Cell and Plant (Exiqon; Denmark). .. RNA concentration was calculated by using the QUBIT fluorometer and the Quant-iT RNA Assay kit (Invitrogen; California, USA).

    Dissection:

    Article Title: Identification and characterization of the doublesex gene of Nasonia
    Article Snippet: Paragraph title: Tissue dissection and RNA purification ... When necessary, RNA was quantified using a Qubit fluorometer (Invitrogen, CA) and a Quant-iT RNA Assay Kit (Invitrogen, CA) or a ND-1000 Spectrophotometer (Nanodrop technologies, UK).

    RNA Extraction:

    Article Title: MicroRNAs in Vitis vinifera cv. Chardonnay Are Differentially Expressed in Response to Diaporthe Species
    Article Snippet: Paragraph title: 2.4. RNA Extraction and Quality Control ... The total RNA yield and quality were measured using a Bioanalyzer 2100 (Agilent Technologies, Palo Alto, CA, USA) using the Agilent RNA 6000 Nano Kit and Modulus™ Single Tube Multimode Reader (Turner Biosystems, Sunnyvale, CA, USA) using the Quant-iT™ RNA Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA).

    Spectrophotometry:

    Article Title: Deficit of Kcnma1 mRNA expression in the dentate gyrus of epileptic rats
    Article Snippet: .. The concentration and purity of total RNA for each sample was determined by the Quant-iT RNA Assay Kit and Q32857 Qubit fluorometer (Carlsbad, Invitrogen, California, USA) and confirmed by optical density measurements at 260 and 280nm using a BioMate 5 UV-visible spectrophotometer (Thermo Spectronic, Waltham, Massachusetts, USA). .. The integrity of the extracted RNA was confirmed by electrophoresis under denaturating condition [ ].

    Article Title: Identification and characterization of the doublesex gene of Nasonia
    Article Snippet: .. When necessary, RNA was quantified using a Qubit fluorometer (Invitrogen, CA) and a Quant-iT RNA Assay Kit (Invitrogen, CA) or a ND-1000 Spectrophotometer (Nanodrop technologies, UK). .. Poly(A) RNA or total RNA were reverse transcribed using SuperScript III (Invitrogen, CA) and Oligo(dT)12-18 primer (Invitrogen, CA) or using RevertAid™ H Minus First Strand cDNA Synthesis Kit (Fermentas, MD) and 3′ RACE adapter supplied with FirstChoice RLM-RACE kit (Ambion, TX).

    Article Title: Semen miRNAs Contained in Exosomes as Non-Invasive Biomarkers for Prostate Cancer Diagnosis
    Article Snippet: RNA concentration was calculated by using the QUBIT fluorometer and the Quant-iT RNA Assay kit (Invitrogen; California, USA). .. All RNA samples presented an OD 260/280 nm ratio ≥ 1,7 when using a Nanodrop UV-Vis spectrophotometer (Thermo Fisher Scientific; Massachusetts, USA).

    Purification:

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: During the process of RNA purification, genomic DNA was removed effectively in a single centrifugation step. .. RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm.

    Article Title: Identification and characterization of the doublesex gene of Nasonia
    Article Snippet: Paragraph title: Tissue dissection and RNA purification ... When necessary, RNA was quantified using a Qubit fluorometer (Invitrogen, CA) and a Quant-iT RNA Assay Kit (Invitrogen, CA) or a ND-1000 Spectrophotometer (Nanodrop technologies, UK).

    Article Title: Identification of the Efflux Transporter of the Fluoroquinolone Antibiotic Ciprofloxacin in Murine Macrophages: Studies with Ciprofloxacin-Resistant Cells ▿
    Article Snippet: .. Total RNA was purified with Turbo DNase (Ambion, Austin, TX), and the concentration was measured with a Qubit fluorimeter, using a Quant-iT RNA assay kit (Invitrogen). cDNA was synthesized using 1 μg of total purified RNA and random hexamer primers (Promega reverse transcription system; Promega Co., Madison, WI). .. Primer pairs for all Mrp (also referred to as Abcc ) genes investigated were designed using Primer 3 and Autoprime programs , taking into account exon-intron boundaries to prevent genomic DNA amplification.

    Real-time Polymerase Chain Reaction:

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: Paragraph title: Real-time PCR ... RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm.

    Concentration Assay:

    Article Title: Deficit of Kcnma1 mRNA expression in the dentate gyrus of epileptic rats
    Article Snippet: .. The concentration and purity of total RNA for each sample was determined by the Quant-iT RNA Assay Kit and Q32857 Qubit fluorometer (Carlsbad, Invitrogen, California, USA) and confirmed by optical density measurements at 260 and 280nm using a BioMate 5 UV-visible spectrophotometer (Thermo Spectronic, Waltham, Massachusetts, USA). .. The integrity of the extracted RNA was confirmed by electrophoresis under denaturating condition [ ].

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: .. RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm. .. First-strand cDNA (cDNA) was synthesized with the iScript™ cDNA synthesis kit (Bio-Rad, Hercules, CA, USA) using 20 μl reaction mixture containing 0.25 μg total RNA, 4 μl 5× iScript reaction mixture, and 1 μl iScript RT.

    Article Title: Identification of the Efflux Transporter of the Fluoroquinolone Antibiotic Ciprofloxacin in Murine Macrophages: Studies with Ciprofloxacin-Resistant Cells ▿
    Article Snippet: .. Total RNA was purified with Turbo DNase (Ambion, Austin, TX), and the concentration was measured with a Qubit fluorimeter, using a Quant-iT RNA assay kit (Invitrogen). cDNA was synthesized using 1 μg of total purified RNA and random hexamer primers (Promega reverse transcription system; Promega Co., Madison, WI). .. Primer pairs for all Mrp (also referred to as Abcc ) genes investigated were designed using Primer 3 and Autoprime programs , taking into account exon-intron boundaries to prevent genomic DNA amplification.

    Article Title: Semen miRNAs Contained in Exosomes as Non-Invasive Biomarkers for Prostate Cancer Diagnosis
    Article Snippet: .. RNA concentration was calculated by using the QUBIT fluorometer and the Quant-iT RNA Assay kit (Invitrogen; California, USA). .. All RNA samples presented an OD 260/280 nm ratio ≥ 1,7 when using a Nanodrop UV-Vis spectrophotometer (Thermo Fisher Scientific; Massachusetts, USA).

    Random Hexamer Labeling:

    Article Title: Identification of the Efflux Transporter of the Fluoroquinolone Antibiotic Ciprofloxacin in Murine Macrophages: Studies with Ciprofloxacin-Resistant Cells ▿
    Article Snippet: .. Total RNA was purified with Turbo DNase (Ambion, Austin, TX), and the concentration was measured with a Qubit fluorimeter, using a Quant-iT RNA assay kit (Invitrogen). cDNA was synthesized using 1 μg of total purified RNA and random hexamer primers (Promega reverse transcription system; Promega Co., Madison, WI). .. Primer pairs for all Mrp (also referred to as Abcc ) genes investigated were designed using Primer 3 and Autoprime programs , taking into account exon-intron boundaries to prevent genomic DNA amplification.

    Polymerase Chain Reaction:

    Article Title: Deficit of Kcnma1 mRNA expression in the dentate gyrus of epileptic rats
    Article Snippet: The concentration and purity of total RNA for each sample was determined by the Quant-iT RNA Assay Kit and Q32857 Qubit fluorometer (Carlsbad, Invitrogen, California, USA) and confirmed by optical density measurements at 260 and 280nm using a BioMate 5 UV-visible spectrophotometer (Thermo Spectronic, Waltham, Massachusetts, USA). .. RT were performed on an iCycler Thermal Cycler PCR System (Bio-Rad Laboratories, Hercules, California, USA), the High Capacity cDNA Reverse Transcription Kit (P/N: 4368814; Applied Biosystems, ABI, California, USA) for synthesis of single-stranded cDNA.

    Article Title: Simvastatin suppresses LPS-induced MMP-1 expression in U937 mononuclear cells by inhibiting protein isoprenylation-mediated ERK activation
    Article Snippet: RNA concentration was determined using a Quant-iT™ RNA assay kit from Invitrogen Corp. RNA purity was determined by the ratio of absorbance readings of samples at 260 and 280 nm. .. The RT reaction mixture was then diluted 1:10 with nuclease-free water and used for PCR amplification of MMP cDNA in the presence of the primers.

    Sequencing:

    Article Title: Integrin-linked kinase regulates smooth muscle differentiation marker gene expression in airway tissue
    Article Snippet: The eluted RNA was quantified using the Quant-iT RNA Assay Kit (Invitrogen). .. Blast was conducted to confirm that primer pairs only matched the sequence of the target genes.

    Quantitation Assay:

    Article Title: Integrin-linked kinase regulates smooth muscle differentiation marker gene expression in airway tissue
    Article Snippet: Paragraph title: Quantitation of mRNA. ... The eluted RNA was quantified using the Quant-iT RNA Assay Kit (Invitrogen).

    Variant Assay:

    Article Title: MicroRNAs in Vitis vinifera cv. Chardonnay Are Differentially Expressed in Response to Diaporthe Species
    Article Snippet: The total RNA yield and quality were measured using a Bioanalyzer 2100 (Agilent Technologies, Palo Alto, CA, USA) using the Agilent RNA 6000 Nano Kit and Modulus™ Single Tube Multimode Reader (Turner Biosystems, Sunnyvale, CA, USA) using the Quant-iT™ RNA Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). .. After RNA quantification, samples were pooled in groups according to the variant of inoculation, resulting in a total of five replicates per variant.

    Software:

    Article Title: Deficit of Kcnma1 mRNA expression in the dentate gyrus of epileptic rats
    Article Snippet: After SE, rats were monitored for detecting of at least two spontaneous seizures (8 h/day) using a JVC MiniDV digital video-camera and researcher-assisted SeizureScan software (Clever Sys., Inc, Reston, Virginia, USA). .. The concentration and purity of total RNA for each sample was determined by the Quant-iT RNA Assay Kit and Q32857 Qubit fluorometer (Carlsbad, Invitrogen, California, USA) and confirmed by optical density measurements at 260 and 280nm using a BioMate 5 UV-visible spectrophotometer (Thermo Spectronic, Waltham, Massachusetts, USA).

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    Thermo Fisher quant it picogreen assay kit
    DNA quantity per scaffold at 3 and 7 days, assessed by <t>PicoGreen</t> assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points
    Quant It Picogreen Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quant it picogreen assay kit/product/Thermo Fisher
    Average 90 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    quant it picogreen assay kit - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    90
    Thermo Fisher quant it ribogreen assay kit
    Polyplexes with different corona chemistries have similar size, zeta potential, and cargo loading but varied stability against high salt concentrations. (A, B) Polyplex siRNA encapsulation efficiency and stability is highest at N + :P − 20. (A) <t>Ribogreen</t> assay reveals polyplex encapsulation plateaus by N + :P − ratio of 10. (B) Polyplexes retain higher stability after a 30 min incubation in 30% FBS at N + :P − 20 compared to N + :P − 10 ( p
    Quant It Ribogreen Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quant it ribogreen assay kit/product/Thermo Fisher
    Average 90 stars, based on 81 article reviews
    Price from $9.99 to $1999.99
    quant it ribogreen assay kit - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    N/A
    The QuantStudio 12K Flex OpenArray Block Rnase P Kit is used for verification of the QuantStudio 12K Flex OpenArray block The kit contains 2 Rnase P arrays plus reagents Each
      Buy from Supplier

    Image Search Results


    DNA quantity per scaffold at 3 and 7 days, assessed by PicoGreen assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points

    Journal: Tissue Engineering and Regenerative Medicine

    Article Title: A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering

    doi: 10.1007/s13770-017-0107-5

    Figure Lengend Snippet: DNA quantity per scaffold at 3 and 7 days, assessed by PicoGreen assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points

    Article Snippet: Total DNA content of the samples was calculated using a Quant-iT™ PicoGreen® assay kit (ThermoFisher, UK) as per the manufacturers’ instructions.

    Techniques: Picogreen Assay

    DNA quantification in DT. ( A ) Nuclei were counted on H and E-stained slides of native tissue and DT and represented as the number of nuclei per µm 2 ; ( B ) Following tissue digestion, DNA was extracted and quantified using the Quant-iT™ PicoGreen ® dsDNA assay. The results are presented as means ± SD. *** p

    Journal: International Journal of Molecular Sciences

    Article Title: Development of a Cytocompatible Scaffold from Pig Immature Testicular Tissue Allowing Human Sertoli Cell Attachment, Proliferation and Functionality

    doi: 10.3390/ijms19010227

    Figure Lengend Snippet: DNA quantification in DT. ( A ) Nuclei were counted on H and E-stained slides of native tissue and DT and represented as the number of nuclei per µm 2 ; ( B ) Following tissue digestion, DNA was extracted and quantified using the Quant-iT™ PicoGreen ® dsDNA assay. The results are presented as means ± SD. *** p

    Article Snippet: For quantification, DNA was extracted using the PureLink™ genomic DNA mini kit (Thermo Fisher, Ghent, Belgium, K1820-01) and quantified using the Quant-iT™ PicoGreen® dsDNA assay kit (Thermo Fisher, P7589) following the manufacturer’s instructions.

    Techniques: Staining, Picogreen Assay

    Polyplexes with different corona chemistries have similar size, zeta potential, and cargo loading but varied stability against high salt concentrations. (A, B) Polyplex siRNA encapsulation efficiency and stability is highest at N + :P − 20. (A) Ribogreen assay reveals polyplex encapsulation plateaus by N + :P − ratio of 10. (B) Polyplexes retain higher stability after a 30 min incubation in 30% FBS at N + :P − 20 compared to N + :P − 10 ( p

    Journal: ACS nano

    Article Title: Zwitterionic Nanocarrier Surface Chemistry Improves siRNA Tumor Delivery and Silencing Activity Relative to Polyethylene Glycol

    doi: 10.1021/acsnano.7b01110

    Figure Lengend Snippet: Polyplexes with different corona chemistries have similar size, zeta potential, and cargo loading but varied stability against high salt concentrations. (A, B) Polyplex siRNA encapsulation efficiency and stability is highest at N + :P − 20. (A) Ribogreen assay reveals polyplex encapsulation plateaus by N + :P − ratio of 10. (B) Polyplexes retain higher stability after a 30 min incubation in 30% FBS at N + :P − 20 compared to N + :P − 10 ( p

    Article Snippet: Polyplex encapsulation efficiency at various N+ :P− ratios was evaluated using a Quant-iT Ribogreen assay kit (ThermoFisher, USA).

    Techniques: Incubation

    Validation of MVP. Transfected cells are expressing MVP-biotin and un-transfected cells were used as negative control. (A) Western blot analysis for the detection of MVP in HEK293F cell lysate and exosomal protein extracts shows that plasmid (MVP-biotin) was successfully expressed in HEK293F cells and was partitioned into exosomes isolated from these transfected cells. ( B ) Protein samples from different stages of MVP purification were separated by SDS-PAGE and the protein bands were detected by Comassie staining. After the pull down assay, the presence of MVP within exosomes was confirmed, i.e. MVP in exosomes from biotinylated MVP-transfected cells PD (pull-down lane), whereas it was absent in exosomes from untransfected cells. The position of the MVP-biotin band is indicated by squares. A representative experiment out of 3 is shown. ( C ) Total amount of RNAs coupled to MVP were extracted and quantified by Quant-iT™ RiboGreen® RNA Assay Kit (ThermoFisher Scientific). Captured RNA was expressed as a percentage of RNA eluted from the beads after the pull-down respect to the total RNA incubated with the beads. Exosomal-MVP was coupled with RNAs and the quantification of RNAs that had co-eluted with MVP showed that the amount of RNA present in the MVP elute (i.e. from MVP-RNA complex) was significantly higher than that from the untransfected control. The experiment was performed in 3-replicates and the graph shows the range of values and median (grey). Average and standard error of three independent experiments are shown. Samples: W = proteins from different wash steps during the MVP purification, and PD = pull-down proteins (MVP) after elution, I = input, and UB = unbound after RBP capture.

    Journal: PLoS ONE

    Article Title: Identification of RNA-binding proteins in exosomes capable of interacting with different types of RNA: RBP-facilitated transport of RNAs into exosomes

    doi: 10.1371/journal.pone.0195969

    Figure Lengend Snippet: Validation of MVP. Transfected cells are expressing MVP-biotin and un-transfected cells were used as negative control. (A) Western blot analysis for the detection of MVP in HEK293F cell lysate and exosomal protein extracts shows that plasmid (MVP-biotin) was successfully expressed in HEK293F cells and was partitioned into exosomes isolated from these transfected cells. ( B ) Protein samples from different stages of MVP purification were separated by SDS-PAGE and the protein bands were detected by Comassie staining. After the pull down assay, the presence of MVP within exosomes was confirmed, i.e. MVP in exosomes from biotinylated MVP-transfected cells PD (pull-down lane), whereas it was absent in exosomes from untransfected cells. The position of the MVP-biotin band is indicated by squares. A representative experiment out of 3 is shown. ( C ) Total amount of RNAs coupled to MVP were extracted and quantified by Quant-iT™ RiboGreen® RNA Assay Kit (ThermoFisher Scientific). Captured RNA was expressed as a percentage of RNA eluted from the beads after the pull-down respect to the total RNA incubated with the beads. Exosomal-MVP was coupled with RNAs and the quantification of RNAs that had co-eluted with MVP showed that the amount of RNA present in the MVP elute (i.e. from MVP-RNA complex) was significantly higher than that from the untransfected control. The experiment was performed in 3-replicates and the graph shows the range of values and median (grey). Average and standard error of three independent experiments are shown. Samples: W = proteins from different wash steps during the MVP purification, and PD = pull-down proteins (MVP) after elution, I = input, and UB = unbound after RBP capture.

    Article Snippet: RNA was quantified by Quant-iT™ RiboGreen® RNA Assay Kit (ThermoFisher Scientific).

    Techniques: Transfection, Expressing, Negative Control, Western Blot, Plasmid Preparation, Isolation, Purification, SDS Page, Staining, Pull Down Assay, Incubation