rneasy kit 74104  (Qiagen)


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    Name:
    RNeasy Plus Kit
    Description:

    Catalog Number:
    74104
    Price:
    None
    Score:
    85
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    Structured Review

    Qiagen rneasy kit 74104

    https://www.bioz.com/result/rneasy kit 74104/product/Qiagen
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    rneasy kit 74104 - by Bioz Stars, 2019-12
    99/100 stars

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    Related Articles

    Transduction:

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: After stripping of phospho-αAMP kinase blots, total AMP kinase (Cell Signaling Technology) immunoreactivity was determined. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. For primary hepatocytes, cells were washed 2× with PBS and lysed with 500 μl of RNeasy lysis buffer supplied with the kit.

    Centrifugation:

    Article Title: Activation of p38 mitogen-activated protein kinase by norepinephrine in T-lineage cells
    Article Snippet: Following cell exposure to agents as indicated in figure legend exposures, cells were collected by centrifugation (200 g , 7 min, RT) and lysed in TRIzol LS Reagent (Invitrogen), at a ratio of 1 ml of reagent to 1 × 107 cells. .. Total cellular RNA was extracted using the RNEasy Mini Prep kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions.

    Amplification:

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells
    Article Snippet: mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. .. mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions.

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen). .. Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen).

    Synthesized:

    Article Title: An N-terminal truncated carboxypeptidase E splice isoform induces tumor growth and is a biomarker for predicting future metastasis in human cancers
    Article Snippet: CPE microarray data were mined from GEO Profile data ( ). .. RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science). .. Semiquantitative PCR was performed to quantify CPE-ΔN transcripts using GoTaq Green Master Mix (Promega).

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Cytometry:

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Quantitative RT-PCR:

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit.

    Article Title: Inhibition of Histone Lysine Methylation Enhances Expression of Cancer-Testis Genes in Lung Cancer Cells: Implications for Adoptive Immunotherapy of Cancer
    Article Snippet: DAC/DP and DAC/TSA treatments were performed as described ( ). .. RNA was isolated using RNeasy mini kit (Qiagen, Valencia, CA). cDNAs were made using reverse transcription kit (Bio-Rad; Hercules, CA). qRT-PCR primers for CT-X genes and β-actin expression are listed in . .. Total cell proteins were extracted and immunoblotting was performed as described ( ) with minor modifications, using primary antibodies listed in , appropriate horseradish peroxidase–conjugated secondary antibodies, and SuperSignal West Pico Chemiluminescent Substrate (Pierce Biotechnology, Rockford, IL).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: Paragraph title: Quantitative RT-PCR ... RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively.

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Two μg of total RNA was used for cDNA synthesis using SuperScript II reverse transcriptase (catalog number 18080-044) and random primers (catalog number 48190-011, Invitrogen) in a 20-μl reaction system, with the following cycles: 25 °C for 5 min, 42 °C for 50 min, 75 °C for 5 min. After reaction, the RT-PCR products were diluted with diethyl pyrocarbonate/H2 O to 100 μl for the real time PCR.

    Real-time Polymerase Chain Reaction:

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title:
    Article Snippet: Paragraph title: RNA Extraction, cDNA Synthesis, and Quantitative Real Time PCR Assay ... Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions.

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. A mouse Th1 and Th2 RT2 -PCR Profiler Array (Qiagen, PAMM-034Z) was used to measure the expression level of 84 T-helper-associated genes in the ABI Prism 7500 Sequence Detection System (ThermoFisher).

    Article Title: Activation of p38 mitogen-activated protein kinase by norepinephrine in T-lineage cells
    Article Snippet: Paragraph title: RNA extraction and quantitative real-time polymerase chain reaction ... Total cellular RNA was extracted using the RNEasy Mini Prep kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions.

    Microarray:

    Article Title: Bcl11b/Ctip2 controls the differentiation of vomeronasal sensory neurons in mice
    Article Snippet: The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany). .. The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany).

    Incubation:

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells
    Article Snippet: mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. .. mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions.

    Luciferase:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. Changes in mRNAs were quantified by optical density measurements. c- fos promoter and AP-1 activities were determined by reporter gene assay as previously described ( ; ; ).

    Activity Assay:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. Changes in mRNAs were quantified by optical density measurements. c- fos promoter and AP-1 activities were determined by reporter gene assay as previously described ( ; ; ).

    Infection:

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Sf9 cells were infected with AcMNPV at a multiplicity of infection (MOI) of 10 TCID50 . .. Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen).

    Expressing:

    Article Title: CXCR3/Ligands Are Significantly Involved in the Tumorigenesis of Basal Cell Carcinomas
    Article Snippet: For gene expression analysis, seven nodular, five superficial, and five morpheiform BCCs were excised and preserved in a RNA stabilization reagent (Qiagen, Mississauga, Ontario) and storage at −80°C before use. .. Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions.

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: Paragraph title: Gene Expression Analysis ... Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit.

    Article Title: Bcl11b/Ctip2 controls the differentiation of vomeronasal sensory neurons in mice
    Article Snippet: The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany). .. The microarray data were linearly normalized with the GAPDH (Probe ID: 1418625_s_at) signal of each preparation using GeneChip operating software (Affymetrix).

    Article Title:
    Article Snippet: Total RNAs were extracted at different times (24, 48, 72, and 96 h) (RNeasy Mini Kit, Qiagen SA), and reverse transcription was performed on total RNA (1 mg) using RevertAid M-MuLV Reverse Transcriptase (GmbH, Germany). .. Total RNAs were extracted at different times (24, 48, 72, and 96 h) (RNeasy Mini Kit, Qiagen SA), and reverse transcription was performed on total RNA (1 mg) using RevertAid M-MuLV Reverse Transcriptase (GmbH, Germany).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: RNA was purified from 1 × 105 cells for gene expression analysis immediately following the thaw of the cryopreserved stocks of cultured cells. .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Article Title: Inhibition of Histone Lysine Methylation Enhances Expression of Cancer-Testis Genes in Lung Cancer Cells: Implications for Adoptive Immunotherapy of Cancer
    Article Snippet: DAC/DP and DAC/TSA treatments were performed as described ( ). .. RNA was isolated using RNeasy mini kit (Qiagen, Valencia, CA). cDNAs were made using reverse transcription kit (Bio-Rad; Hercules, CA). qRT-PCR primers for CT-X genes and β-actin expression are listed in . .. Total cell proteins were extracted and immunoblotting was performed as described ( ) with minor modifications, using primary antibodies listed in , appropriate horseradish peroxidase–conjugated secondary antibodies, and SuperSignal West Pico Chemiluminescent Substrate (Pierce Biotechnology, Rockford, IL).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively.

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: The expression of cytokines involved in Th1 and Th2 responses was evaluated in the lung tissue of immunized mice post-challenge. .. Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401).

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Derivative Assay:

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: The number of passages of the RAC and SAC were closely matched for the populations derived from the 13M (passage 7 for RAC and passage 6 for SAC), 57M (passage 6 for both populations), and 70M donor tissues (passage 3 for both populations). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Transfection:

    Article Title:
    Article Snippet: At 80% confluence, cells were transfected with 40 n m siRNA in endothelial basal medium using HiPerFect transfection agent (Qiagen SA). .. Total RNAs were extracted at different times (24, 48, 72, and 96 h) (RNeasy Mini Kit, Qiagen SA), and reverse transcription was performed on total RNA (1 mg) using RevertAid M-MuLV Reverse Transcriptase (GmbH, Germany).

    Rapid Amplification of cDNA Ends:

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Paragraph title: Total RNA preparation, RT-PCR, and 5′RACE (5′ rapid amplification of cDNA ends) analysis. ... Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen).

    SYBR Green Assay:

    Article Title: Triheptanoin: long-term effects in the very long-chain acyl-CoA dehydrogenase-deficient mouse
    Article Snippet: Total RNA from heart was isolated with the RNeasy mini kit (Qiagen, Hilden, Germany). .. Total RNA from heart was isolated with the RNeasy mini kit (Qiagen, Hilden, Germany).

    Article Title: Activation of p38 mitogen-activated protein kinase by norepinephrine in T-lineage cells
    Article Snippet: Total cellular RNA was extracted using the RNEasy Mini Prep kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions. .. Total cellular RNA was extracted using the RNEasy Mini Prep kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions.

    Northern Blot:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Briefly, cells were separated from the medium and pyruvate-dependent oxidation of NADH was monitored and expressed as micromoles of NADH oxidized per minute per milligram protein. .. Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. Changes in mRNAs were quantified by optical density measurements. c- fos promoter and AP-1 activities were determined by reporter gene assay as previously described ( ; ; ).

    Cell Culture:

    Article Title: Profilin-1 overexpression inhibits proliferation of MDA-MB-231 breast cancer cells partly through p27kip1 upregulation
    Article Snippet: SiRNA transfection was done at a working concentration of 25 nM using Dharmacon transfection reagent according to the manufacturer’s instructions. .. Total RNA was extracted from cells cultured in normal serum-containing growth medium using the RNeasy mini kit commercially available through Qiagen (Valencia, CA). .. RT-PCR reactions were performed using a commercial kit also available through Qiagen.

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: The PC013 and CAF039 cell lines were cocultured or cultured separately for 5 days in R10 culture medium. .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: RNA was purified from 1 × 105 cells for gene expression analysis immediately following the thaw of the cryopreserved stocks of cultured cells. .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Triheptanoin: long-term effects in the very long-chain acyl-CoA dehydrogenase-deficient mouse
    Article Snippet: Paragraph title: RT-PCR analysis ... Total RNA from heart was isolated with the RNeasy mini kit (Qiagen, Hilden, Germany).

    Article Title: CXCR3/Ligands Are Significantly Involved in the Tumorigenesis of Basal Cell Carcinomas
    Article Snippet: Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. .. Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions.

    Article Title: Profilin-1 overexpression inhibits proliferation of MDA-MB-231 breast cancer cells partly through p27kip1 upregulation
    Article Snippet: Paragraph title: RT-PCR ... Total RNA was extracted from cells cultured in normal serum-containing growth medium using the RNeasy mini kit commercially available through Qiagen (Valencia, CA).

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: Paragraph title: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) ... Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR.

    Article Title: Reprogramming of skeletal myoblasts for induction of pluripotency for tumor free cardiomyogenesis in the infarcted heart
    Article Snippet: Paragraph title: Reverse transcription polymerase chain reaction (RT-PCR) ... Isolation of total RNA, and their subsequent first-strand cDNA synthesis, wasperformed using an RNeasy mini kit (Qiagen, Valencia, CA) and an Omniscript Reverse Transcription kit (Qiagen, Valencia, CA), respectively, per the instructions of manufacturer and as described earlier.

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Paragraph title: Total RNA preparation, RT-PCR, and 5′RACE (5′ rapid amplification of cDNA ends) analysis. ... Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen).

    Generated:

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. Five housekeeping genes (HKGs) and proprietary controls were tested in parallel for normalization of the assay results, monitoring genomic DNA contamination, cDNA synthesis and real-time PCR efficiency.

    DNA Sequencing:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Sequencing:

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Real time RT-PCR was performed on 384-well plates in a 20-μl reaction system containing 2 μl of diluted cDNA, 1 μl of primer mixture, 7 μl of H2 O, and 10 μl of TaqMan 2× in reaction mixture.

    Injection:

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Recombinant:

    Article Title: CXCR3/Ligands Are Significantly Involved in the Tumorigenesis of Basal Cell Carcinomas
    Article Snippet: Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. .. The synthesis of cDNA was performed using the SuperScript First Strand Synthesis System for RT-PCR kit (Invitrogen).

    Fluorescence:

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Real time RT-PCR was performed on 384-well plates in a 20-μl reaction system containing 2 μl of diluted cDNA, 1 μl of primer mixture, 7 μl of H2 O, and 10 μl of TaqMan 2× in reaction mixture.

    Mutagenesis:

    Article Title: Lipopolysaccharides mediate leukotoxin secretion in Aggregatibacter actinomycetemcomitans
    Article Snippet: After a two-day growth on TSBYE plates, one colony of the wild-type strain (VT1169) and the rmlC mutant were transferred into 8 ml of freshly prepared TSBYE broth, with or without spectinomycin, and grown overnight. .. Total RNA was purified from the lysate using an RNeasy Mini Kit (QIAGEN Inc., Valencia, CA).

    Isolation:

    Article Title: Triheptanoin: long-term effects in the very long-chain acyl-CoA dehydrogenase-deficient mouse
    Article Snippet: Signals were detected and quantified Fusion FX Analyzer (Peqlab, Erlangen, Germany). .. Total RNA from heart was isolated with the RNeasy mini kit (Qiagen, Hilden, Germany). .. Forward and reverse primers were designed with the Primer Design Tool from NCBI ( ).

    Article Title: CXCR3/Ligands Are Significantly Involved in the Tumorigenesis of Basal Cell Carcinomas
    Article Snippet: Samples were removed from the stabilization reagent, and homogenized using a QIAshredder kit (Qiagen) according to the manufacturer’s instructions for total RNA isolation. .. Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. .. DNase digestion was conducted using RNase-Free DNase Set (Qiagen) after the first buffer washing step of the RNA-bound RNeasy membrane.

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: After stripping of phospho-αAMP kinase blots, total AMP kinase (Cell Signaling Technology) immunoreactivity was determined. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. For primary hepatocytes, cells were washed 2× with PBS and lysed with 500 μl of RNeasy lysis buffer supplied with the kit.

    Article Title: Lipopolysaccharides mediate leukotoxin secretion in Aggregatibacter actinomycetemcomitans
    Article Snippet: Paragraph title: Isolation of total RNA from A. actinomycetemcomitans ... Total RNA was purified from the lysate using an RNeasy Mini Kit (QIAGEN Inc., Valencia, CA).

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: To follow the course of embryonic development, a time-course analysis for multilineage differentiation was performed. hESCs were randomly differentiated as described in , and samples were collected for day 0, 3, 6, 9, 12, 15, 18 and 21 for RT-PCR analysis. .. Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. Primer design was performed using Primer3 ( ).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: The number of passages of the RAC and SAC were closely matched for the populations derived from the 13M (passage 7 for RAC and passage 6 for SAC), 57M (passage 6 for both populations), and 70M donor tissues (passage 3 for both populations). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA). .. Gene expression was measured using real-time quantitative PCR (see Supplementary Materials and Methods for detailed methods).

    Article Title: Inhibition of Histone Lysine Methylation Enhances Expression of Cancer-Testis Genes in Lung Cancer Cells: Implications for Adoptive Immunotherapy of Cancer
    Article Snippet: DAC/DP and DAC/TSA treatments were performed as described ( ). .. RNA was isolated using RNeasy mini kit (Qiagen, Valencia, CA). cDNAs were made using reverse transcription kit (Bio-Rad; Hercules, CA). qRT-PCR primers for CT-X genes and β-actin expression are listed in . .. Total cell proteins were extracted and immunoblotting was performed as described ( ) with minor modifications, using primary antibodies listed in , appropriate horseradish peroxidase–conjugated secondary antibodies, and SuperSignal West Pico Chemiluminescent Substrate (Pierce Biotechnology, Rockford, IL).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was isolated from synaptosomal homogenates by addition of 10% BCP and standard phase separation, followed by overnight precipitation with isopropanol at -20°C. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively.

    Article Title: Reprogramming of skeletal myoblasts for induction of pluripotency for tumor free cardiomyogenesis in the infarcted heart
    Article Snippet: The murine SiPS clone Raf1 was expanded on mitotically inactivated murine embryonic fibroblasts (MEFs; 5×104 cells/cm2 ) as described in . .. Isolation of total RNA, and their subsequent first-strand cDNA synthesis, wasperformed using an RNeasy mini kit (Qiagen, Valencia, CA) and an Omniscript Reverse Transcription kit (Qiagen, Valencia, CA), respectively, per the instructions of manufacturer and as described earlier. .. The primer sequences used are given in .

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: The expression of cytokines involved in Th1 and Th2 responses was evaluated in the lung tissue of immunized mice post-challenge. .. Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. A mouse Th1 and Th2 RT2 -PCR Profiler Array (Qiagen, PAMM-034Z) was used to measure the expression level of 84 T-helper-associated genes in the ABI Prism 7500 Sequence Detection System (ThermoFisher).

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Sf9 cells were infected with AcMNPV at a multiplicity of infection (MOI) of 10 TCID50 . .. Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen). .. Reverse transcription-PCR (RT-PCR) was performed with an RNA PCR kit (v3.0; TaKaRa) using 2.0 μg of total RNA as the template for each time point.

    Subcloning:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Negative Control:

    Article Title:
    Article Snippet: Small interfering RNAs for cathepsin L (siCatL), cathepsin S (siCatS), cathepsin B (siCatB), and a negative control siRNA (siCTL) were obtained from Qiagen SA (Courtaboeuf, France) ( ). .. Total RNAs were extracted at different times (24, 48, 72, and 96 h) (RNeasy Mini Kit, Qiagen SA), and reverse transcription was performed on total RNA (1 mg) using RevertAid M-MuLV Reverse Transcriptase (GmbH, Germany).

    Flow Cytometry:

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Labeling:

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Mouse Assay:

    Article Title: Bcl11b/Ctip2 controls the differentiation of vomeronasal sensory neurons in mice
    Article Snippet: VNOs were obtained from Bcl11b−/− and wild-type mice at P0 and stored in RNA later (Ambion, Austin, TX). .. The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany).

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: The expression of cytokines involved in Th1 and Th2 responses was evaluated in the lung tissue of immunized mice post-challenge. .. Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401).

    Polymerase Chain Reaction:

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit.

    Article Title: An N-terminal truncated carboxypeptidase E splice isoform induces tumor growth and is a biomarker for predicting future metastasis in human cancers
    Article Snippet: Paragraph title: Semiquantitative PCR of WT CPE and CPE-ΔN transcripts in HCC cells. ... RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science).

    Article Title: Profilin-1 overexpression inhibits proliferation of MDA-MB-231 breast cancer cells partly through p27kip1 upregulation
    Article Snippet: Total RNA was extracted from cells cultured in normal serum-containing growth medium using the RNeasy mini kit commercially available through Qiagen (Valencia, CA). .. Total RNA was extracted from cells cultured in normal serum-containing growth medium using the RNeasy mini kit commercially available through Qiagen (Valencia, CA).

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: To follow the course of embryonic development, a time-course analysis for multilineage differentiation was performed. hESCs were randomly differentiated as described in , and samples were collected for day 0, 3, 6, 9, 12, 15, 18 and 21 for RT-PCR analysis. .. Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. Primer design was performed using Primer3 ( ).

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells
    Article Snippet: mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. .. mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions.

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Real time RT-PCR was performed on 384-well plates in a 20-μl reaction system containing 2 μl of diluted cDNA, 1 μl of primer mixture, 7 μl of H2 O, and 10 μl of TaqMan 2× in reaction mixture.

    Construct:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Purification:

    Article Title: Lipopolysaccharides mediate leukotoxin secretion in Aggregatibacter actinomycetemcomitans
    Article Snippet: The cells were centrifuged at 5000 ×g for 10 min and resuspended in 100 µl TE buffer (10 mM Tris-Cl, 1 mM EDTA, pH 8.0) containing 1 mg/ml of lysozyme Type VI (MP Biomedicals, Aurora, OH). .. Total RNA was purified from the lysate using an RNeasy Mini Kit (QIAGEN Inc., Valencia, CA). .. The purified total RNA was pretreated with deoxyribonuclease I (DNase), amplification grade (Invitrogen, Carlsbad, CA), to remove contaminating DNA.

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: RNA was purified from 1 × 105 cells for gene expression analysis immediately following the thaw of the cryopreserved stocks of cultured cells. .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was isolated from synaptosomal homogenates by addition of 10% BCP and standard phase separation, followed by overnight precipitation with isopropanol at -20°C. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen). .. The ac93 -specific primers Ac93P1 (5′-ATGGCGACTAGCAAAACGATCG-3′) and Ac93P2 (5′-TTAATTTACAATTTCAATTCC-3′) were used for PCR amplification to detect the transcription of ac93 .

    Chromatin Immunoprecipitation:

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was isolated from synaptosomal homogenates by addition of 10% BCP and standard phase separation, followed by overnight precipitation with isopropanol at -20°C. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Plasmid Preparation:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Software:

    Article Title: Bcl11b/Ctip2 controls the differentiation of vomeronasal sensory neurons in mice
    Article Snippet: The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany). .. To ensure reproducibility, microarray analyses were performed with RNA samples from six Bcl11b−/− and five wild-type mice.

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Gene expression assays included Mtap2 (MAP2, Rn00565046_m1), Dbn1 (drebrin, Rn00578869_m1), Rtn4 (Nogo-A, Rn00582903_m1), Dlgh4 (PSD-95, Rn00571479_m1), Ywhaq (14-3-3θ, Rn00820722_g1) and Camk2 (CaMKIIα, Rn00563883_m1).

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. Five housekeeping genes (HKGs) and proprietary controls were tested in parallel for normalization of the assay results, monitoring genomic DNA contamination, cDNA synthesis and real-time PCR efficiency.

    Electrophoresis:

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. A thermocycler (Bio-Rad, Munich, Germany) was used for PCR reaction, with the following conditions: denaturation at 95°C/2 min, 22 cycles for β-ACTIN and 35 cycles for target genes, of 95°C/30 s denaturation, 60°C/30 s annealing 72°C/60 s of elongation.

    RNA Extraction:

    Article Title:
    Article Snippet: Paragraph title: RNA Extraction, cDNA Synthesis, and Quantitative Real Time PCR Assay ... Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions.

    Article Title: Activation of p38 mitogen-activated protein kinase by norepinephrine in T-lineage cells
    Article Snippet: Paragraph title: RNA extraction and quantitative real-time polymerase chain reaction ... Total cellular RNA was extracted using the RNEasy Mini Prep kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions.

    shRNA:

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: After stripping of phospho-αAMP kinase blots, total AMP kinase (Cell Signaling Technology) immunoreactivity was determined. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. For primary hepatocytes, cells were washed 2× with PBS and lysed with 500 μl of RNeasy lysis buffer supplied with the kit.

    Concentration Assay:

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR.

    Staining:

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. The double-stranded cDNA was purified using the Sample Cleanup Module and cDNA Cleanup Spin Columns (Affymetrix) and biotin-labeled antisense complementary RNA was synthesized using GeneChip IVT labeling kit (Affymetrix).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Variant Assay:

    Article Title: An N-terminal truncated carboxypeptidase E splice isoform induces tumor growth and is a biomarker for predicting future metastasis in human cancers
    Article Snippet: RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science). .. RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science).

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    Qiagen hmec 1 cells
    Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved <t>HMEC-1</t> cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P
    Hmec 1 Cells, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved HMEC-1 cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved HMEC-1 cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Incubation, Expressing, Western Blot

    Chemerin increases endothelial cell adhesion molecules mRNA expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 4 hours. Real-time quantitative RT-PCR analyses showed that mRNA expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly up-regulated by chemerin in a concentration dependent manner at 4 hours (Figures 2A-2C : *** P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin increases endothelial cell adhesion molecules mRNA expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 4 hours. Real-time quantitative RT-PCR analyses showed that mRNA expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly up-regulated by chemerin in a concentration dependent manner at 4 hours (Figures 2A-2C : *** P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Expressing, Quantitative RT-PCR, Concentration Assay

    Chemerin increases endothelial cell adhesion molecules protein expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 and 24 hours. Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly increased by chemerin in a concentration dependent manner at 12 hours (Figures 3A-3C : * P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin increases endothelial cell adhesion molecules protein expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 and 24 hours. Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly increased by chemerin in a concentration dependent manner at 12 hours (Figures 3A-3C : * P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Expressing, Western Blot, Concentration Assay

    Chemerin increases endothelial cell adhesion molecules protein secretion in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 hours. Densitometric analysis of western blots (conditioned media) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that secretion of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly elevated by chemerin in a concentration dependent manner at 12 hours (Figures 4A-4C : * P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin increases endothelial cell adhesion molecules protein secretion in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 hours. Densitometric analysis of western blots (conditioned media) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that secretion of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly elevated by chemerin in a concentration dependent manner at 12 hours (Figures 4A-4C : * P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Western Blot, Concentration Assay

    Chemerin stimulates monocyte-endothelial cell adhesion via NF-ĸB, MAPK and PI3K/Akt pathways Serum starved chemerin (0-10nM) treated HMEC-1 cells were co-cultured with THP-1 cells for 2 hours. Chemerin enhanced monocyte-endothelial adhesion in a concentration dependent fashion (Figure 6A : * P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin stimulates monocyte-endothelial cell adhesion via NF-ĸB, MAPK and PI3K/Akt pathways Serum starved chemerin (0-10nM) treated HMEC-1 cells were co-cultured with THP-1 cells for 2 hours. Chemerin enhanced monocyte-endothelial adhesion in a concentration dependent fashion (Figure 6A : * P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Cell Culture, Concentration Assay

    Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation ( ** P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation ( ** P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Activation Assay, Stable Transfection, Transfection, Luciferase, Concentration Assay, Activity Assay, Incubation

    Stimulation of an IFN-mediated antiviral response in human transformed or tumor cells upon Poly(I:C) transfection. ( A ) HEK293, HEK293T, NB324K and Hela cultures were transfected with 2 µg/ml of synthetic dsRNA Poly(I:C) (pI:C) or a 150 mM NaCl solution as control, using lipofectamine 2000 for the period indicated in the figure. The respective culture media were then collected, centrifuged to discard cellular debris, and analyzed by Enzyme-linked Immuno-Sorbent Assay (ELISA) for their content in human IFN-β. Each result is represented as mean+standard deviation of three independent experiments. ( B ) Expression of IFN-α and IFN-β transcripts in mock-treated or pI:C-transfected human cell lines was assessed by RT-PCRs. At the indicated time points, total RNAs were extracted from the respective cultures using the RNeasy kit. One µg of RNA was then treated by DNase I in order to remove potential contaminating DNA and reverse transcribed into cDNA. A fraction of the obtained cDNA (1/10) was then analyzed by PCR for its content in type-I IFN transcripts using specific primer pairs. Transcripts encoding the Human 18S ribosomal protein were used as internal loading controls. Data shown are representative of 3 experiments which gave similar results. ( C ) Assessment of the IFN-signaling (Jak/STAT) pathway activation in HEK293, HEK293T, NB324K and Hela cells upon pI:C transfection. At the time indicated mock-treated or pI:C-transfected (2 µg/ml) cultures were harvested by scraping in PBS and centrifuged. Cell pellets were then re-suspended in complete Ripa buffer supplemented with phosphatase and protease inhibitors. Total proteins were extracted from each sample as described in Materials and Methods . Fifty µg total proteins per sample were then subjected to bipartite 8/10% SDS-PAGE, transferred onto membranes, and probed with antibodies specific for phosphorylated and total isoforms of STAT 1 and STAT 2 as well as with an antibody specific to PKR. Actin was used as an internal loading control. Each presented blot is representative of 3 additional which gave similar results.

    Journal: PLoS ONE

    Article Title: TLR-9 Contributes to the Antiviral Innate Immune Sensing of Rodent Parvoviruses MVMp and H-1PV by Normal Human Immune Cells

    doi: 10.1371/journal.pone.0055086

    Figure Lengend Snippet: Stimulation of an IFN-mediated antiviral response in human transformed or tumor cells upon Poly(I:C) transfection. ( A ) HEK293, HEK293T, NB324K and Hela cultures were transfected with 2 µg/ml of synthetic dsRNA Poly(I:C) (pI:C) or a 150 mM NaCl solution as control, using lipofectamine 2000 for the period indicated in the figure. The respective culture media were then collected, centrifuged to discard cellular debris, and analyzed by Enzyme-linked Immuno-Sorbent Assay (ELISA) for their content in human IFN-β. Each result is represented as mean+standard deviation of three independent experiments. ( B ) Expression of IFN-α and IFN-β transcripts in mock-treated or pI:C-transfected human cell lines was assessed by RT-PCRs. At the indicated time points, total RNAs were extracted from the respective cultures using the RNeasy kit. One µg of RNA was then treated by DNase I in order to remove potential contaminating DNA and reverse transcribed into cDNA. A fraction of the obtained cDNA (1/10) was then analyzed by PCR for its content in type-I IFN transcripts using specific primer pairs. Transcripts encoding the Human 18S ribosomal protein were used as internal loading controls. Data shown are representative of 3 experiments which gave similar results. ( C ) Assessment of the IFN-signaling (Jak/STAT) pathway activation in HEK293, HEK293T, NB324K and Hela cells upon pI:C transfection. At the time indicated mock-treated or pI:C-transfected (2 µg/ml) cultures were harvested by scraping in PBS and centrifuged. Cell pellets were then re-suspended in complete Ripa buffer supplemented with phosphatase and protease inhibitors. Total proteins were extracted from each sample as described in Materials and Methods . Fifty µg total proteins per sample were then subjected to bipartite 8/10% SDS-PAGE, transferred onto membranes, and probed with antibodies specific for phosphorylated and total isoforms of STAT 1 and STAT 2 as well as with an antibody specific to PKR. Actin was used as an internal loading control. Each presented blot is representative of 3 additional which gave similar results.

    Article Snippet: Briefly, Total RNAs of mock-treated, parvovirus- or NDV-infected, and/or PolyI:C transfected cells were isolated using an RNeasy mini kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions.

    Techniques: Transformation Assay, Transfection, Enzyme-linked Immunosorbent Assay, Standard Deviation, Expressing, Polymerase Chain Reaction, Activation Assay, SDS Page

    Time-dependent transcriptional up-regulation of type-I IFN encoding mRNAs in MVMp- or H-1PV-infected human cell lines. ( A ) HEK293 and HEK293T, ( B ) NB324K and ( C ) Hela cells were mock-treated for 24 hrs, infected with parvoviruses (5 PFUs/cell) for the indicated times, or infected with NDV (6 HU/10 6 cells) or transfected with pI:C (2 µg/ml) for 15 hrs. At the indicated times, cells were harvested and total RNAs were extracted using the RNeasy kit. One µg was then reverse transcribed into cDNA and 10% of this product was subjected to PCR reactions using sets of primers specific to each indicated cytokine mRNA or viral transcripts. PCR product of 18S ribosomal RNA was used as housekeeping gene to normalize loading. No signal was detected in the samples when omitting the reverse transcriptase. Presented data are representative of 3 experiments which all gave similar results.

    Journal: PLoS ONE

    Article Title: TLR-9 Contributes to the Antiviral Innate Immune Sensing of Rodent Parvoviruses MVMp and H-1PV by Normal Human Immune Cells

    doi: 10.1371/journal.pone.0055086

    Figure Lengend Snippet: Time-dependent transcriptional up-regulation of type-I IFN encoding mRNAs in MVMp- or H-1PV-infected human cell lines. ( A ) HEK293 and HEK293T, ( B ) NB324K and ( C ) Hela cells were mock-treated for 24 hrs, infected with parvoviruses (5 PFUs/cell) for the indicated times, or infected with NDV (6 HU/10 6 cells) or transfected with pI:C (2 µg/ml) for 15 hrs. At the indicated times, cells were harvested and total RNAs were extracted using the RNeasy kit. One µg was then reverse transcribed into cDNA and 10% of this product was subjected to PCR reactions using sets of primers specific to each indicated cytokine mRNA or viral transcripts. PCR product of 18S ribosomal RNA was used as housekeeping gene to normalize loading. No signal was detected in the samples when omitting the reverse transcriptase. Presented data are representative of 3 experiments which all gave similar results.

    Article Snippet: Briefly, Total RNAs of mock-treated, parvovirus- or NDV-infected, and/or PolyI:C transfected cells were isolated using an RNeasy mini kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions.

    Techniques: Infection, Transfection, Polymerase Chain Reaction

    Activation of both IFN-producing and IFN-signaling pathways in hPBMCs upon parvovirus infection and comparison of their intensity to that triggered by NDV. ( A , B and D ) hPBMCs collected from the blood of healthy donors were distributed into 6-well plates at 1×10 7 cells/5 ml culture medium/well. They were then mock-treated or infected with MVMp or H-1PV at 20 PFUs/cell or with NDV (6 HU/10 6 cells). Cultures were harvested in PBS and centrifuged at the time p.i. indicated in each figure. ( A , D ) One half of each pellet was resuspended in Ripa buffer supplemented with phosphatase and protease inhibitors in order to perform Western blot experiments whereas ( B ) total RNAs were extracted from the rest of each cell pellet using the RNeasy kit. ( A , D ) Total proteins were extracted from each sample as described in Materials and Methods . Seventy µg total proteins per sample were then subjected to 10% SDS-PAGE, transferred onto membranes, and probed with antibodies specific for total and phosphorylated STAT 2 and STAT 1 polypeptides as well as for PKR. Actin was used as an internal loading control. Each presented blot is representative of 4 additional which gave similar results. ( B ). One µg of isolated total RNA was then reverse transcribed into cDNA and 10% of this product was subjected to PCR reactions using sets of primers specific to each indicated mRNA. PCR product of 18S ribosomal RNA was used as housekeeping gene to normalize loading. No signal was detected in the samples when omitting the reverse transcriptase. Presented data are representative of 4 experiments which all gave similar results. ( C ) hPBMCs collected from the blood of healthy donors were distributed into 24-well plates at 1×10 6 cells/500 µl culture medium/well. They were then mock-treated or infected with MVMp, H-1PV (20 PFUs/cell) or NDV (6 HU/10 6 cells). After a period of incubation of 24 hrs media were collected, centrifuged in order to discard cellular debris and analyzed by Enzyme-linked Immuno-Sorbent Assay (ELISA) for their content in IFN-α and IFN-β. Results are expressed as means+standard deviations of seven independent experiments.

    Journal: PLoS ONE

    Article Title: TLR-9 Contributes to the Antiviral Innate Immune Sensing of Rodent Parvoviruses MVMp and H-1PV by Normal Human Immune Cells

    doi: 10.1371/journal.pone.0055086

    Figure Lengend Snippet: Activation of both IFN-producing and IFN-signaling pathways in hPBMCs upon parvovirus infection and comparison of their intensity to that triggered by NDV. ( A , B and D ) hPBMCs collected from the blood of healthy donors were distributed into 6-well plates at 1×10 7 cells/5 ml culture medium/well. They were then mock-treated or infected with MVMp or H-1PV at 20 PFUs/cell or with NDV (6 HU/10 6 cells). Cultures were harvested in PBS and centrifuged at the time p.i. indicated in each figure. ( A , D ) One half of each pellet was resuspended in Ripa buffer supplemented with phosphatase and protease inhibitors in order to perform Western blot experiments whereas ( B ) total RNAs were extracted from the rest of each cell pellet using the RNeasy kit. ( A , D ) Total proteins were extracted from each sample as described in Materials and Methods . Seventy µg total proteins per sample were then subjected to 10% SDS-PAGE, transferred onto membranes, and probed with antibodies specific for total and phosphorylated STAT 2 and STAT 1 polypeptides as well as for PKR. Actin was used as an internal loading control. Each presented blot is representative of 4 additional which gave similar results. ( B ). One µg of isolated total RNA was then reverse transcribed into cDNA and 10% of this product was subjected to PCR reactions using sets of primers specific to each indicated mRNA. PCR product of 18S ribosomal RNA was used as housekeeping gene to normalize loading. No signal was detected in the samples when omitting the reverse transcriptase. Presented data are representative of 4 experiments which all gave similar results. ( C ) hPBMCs collected from the blood of healthy donors were distributed into 24-well plates at 1×10 6 cells/500 µl culture medium/well. They were then mock-treated or infected with MVMp, H-1PV (20 PFUs/cell) or NDV (6 HU/10 6 cells). After a period of incubation of 24 hrs media were collected, centrifuged in order to discard cellular debris and analyzed by Enzyme-linked Immuno-Sorbent Assay (ELISA) for their content in IFN-α and IFN-β. Results are expressed as means+standard deviations of seven independent experiments.

    Article Snippet: Briefly, Total RNAs of mock-treated, parvovirus- or NDV-infected, and/or PolyI:C transfected cells were isolated using an RNeasy mini kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions.

    Techniques: Activation Assay, Infection, Western Blot, SDS Page, Isolation, Polymerase Chain Reaction, Incubation, Enzyme-linked Immunosorbent Assay

    Activation of a TLR-9 dependent production of type-I IFNs in parvovirus-infected human Namalwa cells. The cells (1.10 6 cells/well of a 24-well plate) were either infected with MVMp or H-1PV (∼80 PFUs/cell equivalent to 1×10 4 virus genomes/cell), stimulated with the TLR-9 agonist ODN 2395 at 1 µM, or mock-treated. ( A ) At the indicated time points culture supernatants were collected from the respective wells and used to determine the quantity of type-I IFNs released in the culture medium by ELISA experiments. Results are expressed as means+standard deviations of three independent experiments. ( B ) Mock-treated, parvovirus-infected or ODN stimulated Namalwa cells were harvested at the indicated time points and total RNAs were extracted using the RNeasy kit as described previously in Figure 1 . Total RNAs extracted from parvovirus-infected (24 hrs p.i. at 80 PFUs/cell) or pI:C-transfected (15 hrs post-transfection with 2 µg dsRNA/ml) NB324K cells were used as positive or negative controls. Presented data are representative of 3 experiments which all gave similar results. ( C ) Total DNA was harvested at the indicated time points from parvovirus-infected (MOI of 80 PFUs/cell) or mock-treated Namalwa or HEK293T cultures to assess by Southern blot experiments as described in Figure 2 the replication of both parvoviruses (dRF, dimmer replicative form; mRF, monomeric replicative form; ssDNA, single-stranded DNA genome). The blot shown is representative of 3 experiments which gave similar results. ( D ) Cell pellets from mock-treated or parvovirus-infected Namalwa cultures were re-suspended in complete Ripa buffer and Western blotting was performed as described in Figure 6 . Actin was used as an internal loading control. Each presented blot is representative of 3 experiments which gave similar results.

    Journal: PLoS ONE

    Article Title: TLR-9 Contributes to the Antiviral Innate Immune Sensing of Rodent Parvoviruses MVMp and H-1PV by Normal Human Immune Cells

    doi: 10.1371/journal.pone.0055086

    Figure Lengend Snippet: Activation of a TLR-9 dependent production of type-I IFNs in parvovirus-infected human Namalwa cells. The cells (1.10 6 cells/well of a 24-well plate) were either infected with MVMp or H-1PV (∼80 PFUs/cell equivalent to 1×10 4 virus genomes/cell), stimulated with the TLR-9 agonist ODN 2395 at 1 µM, or mock-treated. ( A ) At the indicated time points culture supernatants were collected from the respective wells and used to determine the quantity of type-I IFNs released in the culture medium by ELISA experiments. Results are expressed as means+standard deviations of three independent experiments. ( B ) Mock-treated, parvovirus-infected or ODN stimulated Namalwa cells were harvested at the indicated time points and total RNAs were extracted using the RNeasy kit as described previously in Figure 1 . Total RNAs extracted from parvovirus-infected (24 hrs p.i. at 80 PFUs/cell) or pI:C-transfected (15 hrs post-transfection with 2 µg dsRNA/ml) NB324K cells were used as positive or negative controls. Presented data are representative of 3 experiments which all gave similar results. ( C ) Total DNA was harvested at the indicated time points from parvovirus-infected (MOI of 80 PFUs/cell) or mock-treated Namalwa or HEK293T cultures to assess by Southern blot experiments as described in Figure 2 the replication of both parvoviruses (dRF, dimmer replicative form; mRF, monomeric replicative form; ssDNA, single-stranded DNA genome). The blot shown is representative of 3 experiments which gave similar results. ( D ) Cell pellets from mock-treated or parvovirus-infected Namalwa cultures were re-suspended in complete Ripa buffer and Western blotting was performed as described in Figure 6 . Actin was used as an internal loading control. Each presented blot is representative of 3 experiments which gave similar results.

    Article Snippet: Briefly, Total RNAs of mock-treated, parvovirus- or NDV-infected, and/or PolyI:C transfected cells were isolated using an RNeasy mini kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions.

    Techniques: Activation Assay, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Southern Blot, Western Blot

    Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Journal: PLoS ONE

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    doi: 10.1371/journal.pone.0196913

    Figure Lengend Snippet: Comparison of sera exosomal RNA using four different RNA extraction methods. (A) Total RNA yield from ultracentrifugation (UC) and ExoQuick (EQ) treated samples using the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and AllPrep DNA/RNA Mini Kit. (B) Demonstration of RNA quality measured by OD 260 /OD 280 in EQ and UC treated samples. Data are shown as the mean ± SD from six independent patient samples. *** P

    Article Snippet: RNA yield and OD260 /OD280 ratios were therefore compared for exosomal RNA isolated using four different RNA extraction methods: the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and the AllPrep DNA/RNA Mini Kit ( ).

    Techniques: RNA Extraction

    Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Journal: PLoS ONE

    Article Title: Optimizing exosomal RNA isolation for RNA-Seq analyses of archival sera specimens

    doi: 10.1371/journal.pone.0196913

    Figure Lengend Snippet: Exosomal RNAs are stable over two decades. Dot plots of total RNA yield and OD 260 /OD 280 from 105 EQ-treated archival patient samples using the RNeasy Mini Kit combined with TRIzol LS. Storage time for each sample is indicated on the x-axis. The Spearman’s rank correlations for RNA yield versus storing time and OD 260 /OD 280 versus storage time are 0.185 ( P = 0.06) and 0.04 ( P = 0.70), respectively.

    Article Snippet: RNA yield and OD260 /OD280 ratios were therefore compared for exosomal RNA isolated using four different RNA extraction methods: the RNeasy Mini Kit combined with TRIzol LS, the RNeasy Mini Kit alone, conventional RNA precipitation, and the AllPrep DNA/RNA Mini Kit ( ).

    Techniques: