salmeterol  (Qiagen)


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    QuantiTect Rev Transcription Kit
    Description:
    For fast cDNA synthesis enabling sensitive real time two step RT PCR for gene expression analysis Kit contents Qiagen QuantiTect Reverse Transcription Kit 10 x 20L rxns RNA Template Sample Two step cDNA Production Genomic DNA Digestion Reaction Type For Fast cDNA Synthesis Enabling Sensitive Real time Two step RT PCR for Gene Expression Analysis Includes 10 x 20L rxns 100L 7x gDNA Wipeout Buffer 10L Quantiscript Reverse Transcriptase 200L 5x Quantiscript RT Buffer 50L RT Primer Mix 1 9mL RNase free Water Benefits cDNA synthesis and gDNA removal in only 20 minutes High cDNA yields even from low abundance transcripts cDNA synthesis from 5 and 3 regions of transcripts No need to design RNA specific primers or prob
    Catalog Number:
    205310
    Price:
    101
    Category:
    QuantiTect Reverse Transcription Kit
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    Structured Review

    Qiagen salmeterol
    QuantiTect Rev Transcription Kit
    For fast cDNA synthesis enabling sensitive real time two step RT PCR for gene expression analysis Kit contents Qiagen QuantiTect Reverse Transcription Kit 10 x 20L rxns RNA Template Sample Two step cDNA Production Genomic DNA Digestion Reaction Type For Fast cDNA Synthesis Enabling Sensitive Real time Two step RT PCR for Gene Expression Analysis Includes 10 x 20L rxns 100L 7x gDNA Wipeout Buffer 10L Quantiscript Reverse Transcriptase 200L 5x Quantiscript RT Buffer 50L RT Primer Mix 1 9mL RNase free Water Benefits cDNA synthesis and gDNA removal in only 20 minutes High cDNA yields even from low abundance transcripts cDNA synthesis from 5 and 3 regions of transcripts No need to design RNA specific primers or prob
    https://www.bioz.com/result/salmeterol/product/Qiagen
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    salmeterol - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "Efficacy of salmeterol and formoterol combination treatment in mice with chronic obstructive pulmonary disease"

    Article Title: Efficacy of salmeterol and formoterol combination treatment in mice with chronic obstructive pulmonary disease

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2017.5562

    Histopathological assessment of lung tissue samples from mice with chronic obstructive pulmonary disease after salmeterol and/or formoterol treatment. Representative images of (A) hypertrophy and hyperplasia of goblet cells on airway surface, (B) alveolar structure, (C) airway smooth muscle, (D) smooth muscle cells in the small arteries and (E) inflammatory cell infiltration and smooth muscle cells. Magnification, ×40.
    Figure Legend Snippet: Histopathological assessment of lung tissue samples from mice with chronic obstructive pulmonary disease after salmeterol and/or formoterol treatment. Representative images of (A) hypertrophy and hyperplasia of goblet cells on airway surface, (B) alveolar structure, (C) airway smooth muscle, (D) smooth muscle cells in the small arteries and (E) inflammatory cell infiltration and smooth muscle cells. Magnification, ×40.

    Techniques Used: Mouse Assay

    Expression of biological markers of chronic obstructive pulmonary disease in experimental mice. (A) KL-6, (B) CCL-18, (C) MMP-7 and (D) SP-A mRNA expression levels were analyzed in mice treated with salmeterol and/or formoterol. **P
    Figure Legend Snippet: Expression of biological markers of chronic obstructive pulmonary disease in experimental mice. (A) KL-6, (B) CCL-18, (C) MMP-7 and (D) SP-A mRNA expression levels were analyzed in mice treated with salmeterol and/or formoterol. **P

    Techniques Used: Expressing, Mouse Assay

    Expression of inflammatory factors in mice with COPD after salmeterol and/or formoterol treatment. (A) IFN-γ, (B) IL-18, (C) IL-4, (D) IL-1, (E) TNF-α and (F) TGF-β mRNA expression in the lung cells of mice with COPD treated with salmeterol and/or formoterol. **P
    Figure Legend Snippet: Expression of inflammatory factors in mice with COPD after salmeterol and/or formoterol treatment. (A) IFN-γ, (B) IL-18, (C) IL-4, (D) IL-1, (E) TNF-α and (F) TGF-β mRNA expression in the lung cells of mice with COPD treated with salmeterol and/or formoterol. **P

    Techniques Used: Expressing, Mouse Assay

    Therapeutic effects of salmeterol and/or formoterol on mice with COPD. (A) Body weight of mice with COPD during treatment. (B) COPD assessment test scores of experimental mice treated with salmeterol and/or formoterol. (C) Functional residual capacity in mice with COPD after salmeterol and/or formoterol treatment. (D) Inspiratory resistance in mice with COPD after salmeterol and/or formoterol treatment. **P
    Figure Legend Snippet: Therapeutic effects of salmeterol and/or formoterol on mice with COPD. (A) Body weight of mice with COPD during treatment. (B) COPD assessment test scores of experimental mice treated with salmeterol and/or formoterol. (C) Functional residual capacity in mice with COPD after salmeterol and/or formoterol treatment. (D) Inspiratory resistance in mice with COPD after salmeterol and/or formoterol treatment. **P

    Techniques Used: Mouse Assay, Functional Assay

    2) Product Images from "Efficacy of salmeterol and formoterol combination treatment in mice with chronic obstructive pulmonary disease"

    Article Title: Efficacy of salmeterol and formoterol combination treatment in mice with chronic obstructive pulmonary disease

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2017.5562

    Histopathological assessment of lung tissue samples from mice with chronic obstructive pulmonary disease after salmeterol and/or formoterol treatment. Representative images of (A) hypertrophy and hyperplasia of goblet cells on airway surface, (B) alveolar structure, (C) airway smooth muscle, (D) smooth muscle cells in the small arteries and (E) inflammatory cell infiltration and smooth muscle cells. Magnification, ×40.
    Figure Legend Snippet: Histopathological assessment of lung tissue samples from mice with chronic obstructive pulmonary disease after salmeterol and/or formoterol treatment. Representative images of (A) hypertrophy and hyperplasia of goblet cells on airway surface, (B) alveolar structure, (C) airway smooth muscle, (D) smooth muscle cells in the small arteries and (E) inflammatory cell infiltration and smooth muscle cells. Magnification, ×40.

    Techniques Used: Mouse Assay

    Expression of biological markers of chronic obstructive pulmonary disease in experimental mice. (A) KL-6, (B) CCL-18, (C) MMP-7 and (D) SP-A mRNA expression levels were analyzed in mice treated with salmeterol and/or formoterol. **P
    Figure Legend Snippet: Expression of biological markers of chronic obstructive pulmonary disease in experimental mice. (A) KL-6, (B) CCL-18, (C) MMP-7 and (D) SP-A mRNA expression levels were analyzed in mice treated with salmeterol and/or formoterol. **P

    Techniques Used: Expressing, Mouse Assay

    Expression of inflammatory factors in mice with COPD after salmeterol and/or formoterol treatment. (A) IFN-γ, (B) IL-18, (C) IL-4, (D) IL-1, (E) TNF-α and (F) TGF-β mRNA expression in the lung cells of mice with COPD treated with salmeterol and/or formoterol. **P
    Figure Legend Snippet: Expression of inflammatory factors in mice with COPD after salmeterol and/or formoterol treatment. (A) IFN-γ, (B) IL-18, (C) IL-4, (D) IL-1, (E) TNF-α and (F) TGF-β mRNA expression in the lung cells of mice with COPD treated with salmeterol and/or formoterol. **P

    Techniques Used: Expressing, Mouse Assay

    Therapeutic effects of salmeterol and/or formoterol on mice with COPD. (A) Body weight of mice with COPD during treatment. (B) COPD assessment test scores of experimental mice treated with salmeterol and/or formoterol. (C) Functional residual capacity in mice with COPD after salmeterol and/or formoterol treatment. (D) Inspiratory resistance in mice with COPD after salmeterol and/or formoterol treatment. **P
    Figure Legend Snippet: Therapeutic effects of salmeterol and/or formoterol on mice with COPD. (A) Body weight of mice with COPD during treatment. (B) COPD assessment test scores of experimental mice treated with salmeterol and/or formoterol. (C) Functional residual capacity in mice with COPD after salmeterol and/or formoterol treatment. (D) Inspiratory resistance in mice with COPD after salmeterol and/or formoterol treatment. **P

    Techniques Used: Mouse Assay, Functional Assay

    3) Product Images from "Mercury Reduction and Methyl Mercury Degradation by the Soil Bacterium Xanthobacter autotrophicus Py2"

    Article Title: Mercury Reduction and Methyl Mercury Degradation by the Soil Bacterium Xanthobacter autotrophicus Py2

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.01982-15

    MICs of Hg(II) and MeHg in Py2. (A) Hg(II) MICs. The 0 and 0.1 μM Hg(II) growth curves are overlapping. (B) MeHg MICs. The 0, 0.02, and 0.05 μM MeHg growth curves overlap. Error bars represent the standard deviations of the means from
    Figure Legend Snippet: MICs of Hg(II) and MeHg in Py2. (A) Hg(II) MICs. The 0 and 0.1 μM Hg(II) growth curves are overlapping. (B) MeHg MICs. The 0, 0.02, and 0.05 μM MeHg growth curves overlap. Error bars represent the standard deviations of the means from

    Techniques Used:

    Loss of Hg(II) in the presence of Py2. (A) Loss of Hg(II) by growing cultures of Py2. (B) Loss of Hg(II) by Py2 suspended in buffer. Pretreated cells were exposed to 0.02 μM Hg(II) in growth media for 1 h prior to the assay. Mock treated cells
    Figure Legend Snippet: Loss of Hg(II) in the presence of Py2. (A) Loss of Hg(II) by growing cultures of Py2. (B) Loss of Hg(II) by Py2 suspended in buffer. Pretreated cells were exposed to 0.02 μM Hg(II) in growth media for 1 h prior to the assay. Mock treated cells

    Techniques Used:

    Gel electrophoresis of reverse transcription-PCR (RT-PCR) products from intragenic regions of the Py2 mer operon. Three independent cultures (1, 2, and 3) were split into two aliquots, one of which (Hg) was exposed to 0.1 μM Hg(II) and one of
    Figure Legend Snippet: Gel electrophoresis of reverse transcription-PCR (RT-PCR) products from intragenic regions of the Py2 mer operon. Three independent cultures (1, 2, and 3) were split into two aliquots, one of which (Hg) was exposed to 0.1 μM Hg(II) and one of

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

    Capture of Hg(0) in Py2 cultures exposed to Hg(II) or MeHg. (A) Data from experiments conducted with Hg(II). (B) Data from experiments conducted with MeHg. Gray bars represent Hg present in the reaction vessel, and white bars represent Hg present in the
    Figure Legend Snippet: Capture of Hg(0) in Py2 cultures exposed to Hg(II) or MeHg. (A) Data from experiments conducted with Hg(II). (B) Data from experiments conducted with MeHg. Gray bars represent Hg present in the reaction vessel, and white bars represent Hg present in the

    Techniques Used:

    Arrangement of potential mer operons in Xanthobacter autotrophicus Py2. R , merR ; T , merT ; P , merP ; A , merA ; K , potential glutathione reductase. Locus tags are indicated above the genes.
    Figure Legend Snippet: Arrangement of potential mer operons in Xanthobacter autotrophicus Py2. R , merR ; T , merT ; P , merP ; A , merA ; K , potential glutathione reductase. Locus tags are indicated above the genes.

    Techniques Used:

    Gel electrophoresis of RT-PCR products from intergenic regions the Py2 mer operon. (A) Intergenic region between merA 1 and merK 1 . (B) Intergenic region between merA 2 and merK 2 . The template cDNA and no-RT samples were identical to the Hg(II)-exposed samples
    Figure Legend Snippet: Gel electrophoresis of RT-PCR products from intergenic regions the Py2 mer operon. (A) Intergenic region between merA 1 and merK 1 . (B) Intergenic region between merA 2 and merK 2 . The template cDNA and no-RT samples were identical to the Hg(II)-exposed samples

    Techniques Used: Nucleic Acid Electrophoresis, Reverse Transcription Polymerase Chain Reaction

    4) Product Images from "Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival, et al. Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival"

    Article Title: Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival, et al. Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival

    Journal: American Journal of Transplantation

    doi: 10.1111/ajt.14765

    RT q PCR analysis showing relative gene expression levels of IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, TNF ‐α, and IFN ‐γ in allograft skin (A – H) and muscle (I – P). For RT q PCR , a total volume of 15 μl per reaction was used containing 50 ng cDNA template, 1.5 μl QuantiTect Primer Assay (Qiagen), 7.5 μl QuantiTect SYBR green PCR kit (Qiagen), and 5 μl ddH2O. Primers for IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, IFN ‐γ, and TNF ‐α were obtained from Qiagen. Cycling conditions included a hot start activation (5 minutes, 95°C), 40 cycles of 10 seconds denaturation (95°C), and annealing and extension (30 seconds, 60°C). Amplicons were quantified with the comparative threshold cycle ( CT ) method; data acquisition was performed using the 7500 System SDS Software Version 2.0.5 (Applied Biosystems). Amplification specificity was checked using melting curve according to the manufacturer's instructions. All samples were measured in duplicate and respective results were averaged. A–P, In allograft skin (A–H) and muscle (I–P) no significant differences in relative gene expression of cytokines was observed between anti‐ IL ‐1β‐treatment groups 3 and 4 and group 2 controls. Expression levels of all markers were normalized to those of group 2
    Figure Legend Snippet: RT q PCR analysis showing relative gene expression levels of IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, TNF ‐α, and IFN ‐γ in allograft skin (A – H) and muscle (I – P). For RT q PCR , a total volume of 15 μl per reaction was used containing 50 ng cDNA template, 1.5 μl QuantiTect Primer Assay (Qiagen), 7.5 μl QuantiTect SYBR green PCR kit (Qiagen), and 5 μl ddH2O. Primers for IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, IFN ‐γ, and TNF ‐α were obtained from Qiagen. Cycling conditions included a hot start activation (5 minutes, 95°C), 40 cycles of 10 seconds denaturation (95°C), and annealing and extension (30 seconds, 60°C). Amplicons were quantified with the comparative threshold cycle ( CT ) method; data acquisition was performed using the 7500 System SDS Software Version 2.0.5 (Applied Biosystems). Amplification specificity was checked using melting curve according to the manufacturer's instructions. All samples were measured in duplicate and respective results were averaged. A–P, In allograft skin (A–H) and muscle (I–P) no significant differences in relative gene expression of cytokines was observed between anti‐ IL ‐1β‐treatment groups 3 and 4 and group 2 controls. Expression levels of all markers were normalized to those of group 2

    Techniques Used: Polymerase Chain Reaction, Expressing, SYBR Green Assay, Activation Assay, Software, Amplification

    5) Product Images from "The distal upstream promoter in Ly49 genes, Pro1, is active in mature NK cells and T cells, does not require TATA boxes, and displays enhancer activity"

    Article Title: The distal upstream promoter in Ly49 genes, Pro1, is active in mature NK cells and T cells, does not require TATA boxes, and displays enhancer activity

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1401450

    Pro1 displays selective activity in NK and T cell lines
    Figure Legend Snippet: Pro1 displays selective activity in NK and T cell lines

    Techniques Used: Activity Assay

    Mapping of key residues required for Pro1 promoter activity
    Figure Legend Snippet: Mapping of key residues required for Pro1 promoter activity

    Techniques Used: Activity Assay

    Pro1 promoter activity does not require TATA boxes
    Figure Legend Snippet: Pro1 promoter activity does not require TATA boxes

    Techniques Used: Activity Assay

    Pro1 displays enhancer activity
    Figure Legend Snippet: Pro1 displays enhancer activity

    Techniques Used: Activity Assay

    RT-PCR analysis of Pro1 and Pro2 transcripts in cell lines
    Figure Legend Snippet: RT-PCR analysis of Pro1 and Pro2 transcripts in cell lines

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    RT-PCR analysis of Pro1 and Pro2 transcripts in fresh cells
    Figure Legend Snippet: RT-PCR analysis of Pro1 and Pro2 transcripts in fresh cells

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    Promoter activity of mouse Pro1-fragments
    Figure Legend Snippet: Promoter activity of mouse Pro1-fragments

    Techniques Used: Activity Assay

    Promoter activity of rat Pro1-fragments
    Figure Legend Snippet: Promoter activity of rat Pro1-fragments

    Techniques Used: Activity Assay

    6) Product Images from "Sox9 regulates cell state and activity of embryonic mouse mammary progenitor cells"

    Article Title: Sox9 regulates cell state and activity of embryonic mouse mammary progenitor cells

    Journal: Communications Biology

    doi: 10.1038/s42003-018-0215-3

    Effect of Sox9 ablation on embryonic mammary progenitor cell fate and function. a qRT-PCR for Sox9 , Zeb1, Snai2 , Twist , Procr , Trp63 , Acta2 , and Ecad in e1/Co#1 and e1/ Sox9 -KO#1 cells. Relative quantification of each sample is normalised to that of e1/Co#1 and shown in log 2 plot with error bar of triplicates ( n = 3, mean ± s.e.m.). b Heatmap from RNA-seq of e1/control and e1/ Sox9 -KO cells of genes that were significantly up- or down-regulated in all three e1/ Sox9 -KO subclones using both DESeq and Intensity Difference test. c In vitro alveologenesis assay results. Formation of alveoli-like structure and milk production was assessed by morphological changes using bright-field microscopy, as well as IF using anti-milk antibody (red) and DAPI. Scale bar, 200 μm. d In vitro alveologenesis assay results. Quantification of number of alveoli
    Figure Legend Snippet: Effect of Sox9 ablation on embryonic mammary progenitor cell fate and function. a qRT-PCR for Sox9 , Zeb1, Snai2 , Twist , Procr , Trp63 , Acta2 , and Ecad in e1/Co#1 and e1/ Sox9 -KO#1 cells. Relative quantification of each sample is normalised to that of e1/Co#1 and shown in log 2 plot with error bar of triplicates ( n = 3, mean ± s.e.m.). b Heatmap from RNA-seq of e1/control and e1/ Sox9 -KO cells of genes that were significantly up- or down-regulated in all three e1/ Sox9 -KO subclones using both DESeq and Intensity Difference test. c In vitro alveologenesis assay results. Formation of alveoli-like structure and milk production was assessed by morphological changes using bright-field microscopy, as well as IF using anti-milk antibody (red) and DAPI. Scale bar, 200 μm. d In vitro alveologenesis assay results. Quantification of number of alveoli

    Techniques Used: Quantitative RT-PCR, RNA Sequencing Assay, In Vitro, Microscopy

    Loss of Sox9 in e1 enhances colony formation ability and alters sphere morphology. a Schematic illustration for establishment of Sox9 -targeted subclones from Cas9-expressing e1. b qRT-PCR for Sox9 and EMT regulator, Zeb1 . Each RQ is normalised to Co#1. Statistical significance between control group and Sox9- knockout group was computed using unpaired t test. ** P ≤ 0.01, * P ≤ 0.05. Two-tailed P values are 0.0306 for Sox9 and 0.0036 for Zeb1 , respectively ( n = 3, mean with 95% confidence interval). c Immunohistochemistry for Sox9 and Zeb1 showing Zeb1 expression is decreased by Sox9 reduction. Scale bar, 200 μm. d Representative images of spheres grown in methylcellulose (left) and colony formation ability presented in box plots (right) with minimum and maximum values showing number of control or Sox9- targeted cells (out of 10,000 cells plated) that gave rise to spheres in anchorage-independent cultures. Statistical significance was calculated using unpaired t test. Two-tailed **** P = 0.0001 ( n = 3, mean ± s.e.m.). Scale bar, 100 μm. e Representative images of spheroids grown in BME from control and Sox9- targeted cells. Scale bar, 400 μm. f Quantification of area of spheroids and g increase in area of protrusions from spheroids grown in BME from e1/control and e1 -Sox9- KO cells ( n = 8, mean + S.D.) Statistical significance was calculated using one-way analysis of variance (ANOVA) and multiple comparisons, *** P = 0.0002 in f and g . e1, embryonic mammary progenitor cell 1; Co, control, non - targeted cells; KO, knockout; Sox9- targeted cells; MEC, mammary epithelial cell
    Figure Legend Snippet: Loss of Sox9 in e1 enhances colony formation ability and alters sphere morphology. a Schematic illustration for establishment of Sox9 -targeted subclones from Cas9-expressing e1. b qRT-PCR for Sox9 and EMT regulator, Zeb1 . Each RQ is normalised to Co#1. Statistical significance between control group and Sox9- knockout group was computed using unpaired t test. ** P ≤ 0.01, * P ≤ 0.05. Two-tailed P values are 0.0306 for Sox9 and 0.0036 for Zeb1 , respectively ( n = 3, mean with 95% confidence interval). c Immunohistochemistry for Sox9 and Zeb1 showing Zeb1 expression is decreased by Sox9 reduction. Scale bar, 200 μm. d Representative images of spheres grown in methylcellulose (left) and colony formation ability presented in box plots (right) with minimum and maximum values showing number of control or Sox9- targeted cells (out of 10,000 cells plated) that gave rise to spheres in anchorage-independent cultures. Statistical significance was calculated using unpaired t test. Two-tailed **** P = 0.0001 ( n = 3, mean ± s.e.m.). Scale bar, 100 μm. e Representative images of spheroids grown in BME from control and Sox9- targeted cells. Scale bar, 400 μm. f Quantification of area of spheroids and g increase in area of protrusions from spheroids grown in BME from e1/control and e1 -Sox9- KO cells ( n = 8, mean + S.D.) Statistical significance was calculated using one-way analysis of variance (ANOVA) and multiple comparisons, *** P = 0.0002 in f and g . e1, embryonic mammary progenitor cell 1; Co, control, non - targeted cells; KO, knockout; Sox9- targeted cells; MEC, mammary epithelial cell

    Techniques Used: Expressing, Quantitative RT-PCR, Knock-Out, Two Tailed Test, Immunohistochemistry

    Progenitor and lineage-associated marker expression in select eMPCs. a Heatmap depicting stem cell and epithelial marker expression in four eMPC selected for further characterisation. b qRT-PCR for Aldh1a1 , Krt19 , Krt18 , Sox9 , Snai2 , Twist1 , Procr , Trp63 , Fabp4 in eMPC clones. Relative quantification (RQ) of each sample is normalised to that of ePool and shown in log 2 plot with error bar of triplicates ( n = 3, mean ± s.e.m.). c Immunohistochemistry staining of Sox9 and Twist expression in eMPC. Scale bar, 200 μm. eMPC, embryonic mammary progenitor cell; MEF, mouse embryonic fibroblast; MSC, mesenchymal stem cell; ESC, embryonic stem cell; e1 and e2, single-cell-derived clones from GFP− cells; eG1 and eG2, single-cell-derived clones from GFP+ cells; ePool, pool of eMPCs; MaSC, mammary stem cell; VEC, vascular endothelial cell; HSC, hematopoietic stem cell
    Figure Legend Snippet: Progenitor and lineage-associated marker expression in select eMPCs. a Heatmap depicting stem cell and epithelial marker expression in four eMPC selected for further characterisation. b qRT-PCR for Aldh1a1 , Krt19 , Krt18 , Sox9 , Snai2 , Twist1 , Procr , Trp63 , Fabp4 in eMPC clones. Relative quantification (RQ) of each sample is normalised to that of ePool and shown in log 2 plot with error bar of triplicates ( n = 3, mean ± s.e.m.). c Immunohistochemistry staining of Sox9 and Twist expression in eMPC. Scale bar, 200 μm. eMPC, embryonic mammary progenitor cell; MEF, mouse embryonic fibroblast; MSC, mesenchymal stem cell; ESC, embryonic stem cell; e1 and e2, single-cell-derived clones from GFP− cells; eG1 and eG2, single-cell-derived clones from GFP+ cells; ePool, pool of eMPCs; MaSC, mammary stem cell; VEC, vascular endothelial cell; HSC, hematopoietic stem cell

    Techniques Used: Marker, Expressing, Quantitative RT-PCR, Clone Assay, Immunohistochemistry, Staining, Derivative Assay

    7) Product Images from "HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ▿ ▿ †"

    Article Title: HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ▿ ▿ †

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00106-10

    RT-PCR analysis to evaluate transcriptional linkage of PG0104 to the 5′ end of the capsule operon and transcription through the hairpin region. Total RNA was isolated from the wild-type strain, and cDNA was generated as described in Materials and Methods with the QuantiTect reverse transcription kit. In the subsequent PCRs, the following primer combinations were used: RT_end104 (lane 1), RT_link104 (lane 2), RT_hairpin (lane 3), RT_link106 (lane 4), RT_start106 (lane 5). The figure demonstrates linkage of PG0104 to the hairpin, transcription of the hairpin, and linkage of the hairpin to PG0106. M, DNA size markers.
    Figure Legend Snippet: RT-PCR analysis to evaluate transcriptional linkage of PG0104 to the 5′ end of the capsule operon and transcription through the hairpin region. Total RNA was isolated from the wild-type strain, and cDNA was generated as described in Materials and Methods with the QuantiTect reverse transcription kit. In the subsequent PCRs, the following primer combinations were used: RT_end104 (lane 1), RT_link104 (lane 2), RT_hairpin (lane 3), RT_link106 (lane 4), RT_start106 (lane 5). The figure demonstrates linkage of PG0104 to the hairpin, transcription of the hairpin, and linkage of the hairpin to PG0106. M, DNA size markers.

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

    8) Product Images from "Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties"

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0145749

    RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .
    Figure Legend Snippet: RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Agarose Gel Electrophoresis

    9) Product Images from "Toll like Receptor 2 engagement on CD4+ T cells promotes TH9 differentiation and function"

    Article Title: Toll like Receptor 2 engagement on CD4+ T cells promotes TH9 differentiation and function

    Journal: European journal of immunology

    doi: 10.1002/eji.201646846

    Effect of TLR2 engagement on the expression of TH9, TH1 and TH2 transcription factors under non-polarizing or TH9− polarizing conditions. Naïve CD4 + T cells were stimulated with anti-CD3 and anti-CD28 mAbs alone (non-polarizing) or combined with TGF-β and IL-4 (TH9− polarizing) and with or without P3CSK4. Batf (A), Pu.1 (B), Tbx21 (C) and Gata3 (D) mRNA expression was determined by RT-PCR and normalized to actin. Bars represent means ± SD of three independent experiments. * p
    Figure Legend Snippet: Effect of TLR2 engagement on the expression of TH9, TH1 and TH2 transcription factors under non-polarizing or TH9− polarizing conditions. Naïve CD4 + T cells were stimulated with anti-CD3 and anti-CD28 mAbs alone (non-polarizing) or combined with TGF-β and IL-4 (TH9− polarizing) and with or without P3CSK4. Batf (A), Pu.1 (B), Tbx21 (C) and Gata3 (D) mRNA expression was determined by RT-PCR and normalized to actin. Bars represent means ± SD of three independent experiments. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    TLR2 engagement on CD4+ T cells enhances IL9 mRNA and protein expression driven by polyclonal activation and TGF-β and IL-4. Naïve CD4+T cells from WT mice were stimulated with anti-CD3 and anti-CD28 mAbs and exogenous TGF-β (5ng/ml) and IL-4 (10ng/ml), with or without P3CSK4 (1μg/ml) for 48h. (A) Relative expression of Il9 mRNA under polarizing conditions with or without P3CSK4 (n=3). (B) Flow cytometric analysis of CD4+ T cells labeled for intracellular IL-9 and IFN-γ. (C) IL-9 in culture supernatants was determined by ELISA. (D) IFN-γ in culture supernatants was determined by ELISA. Means ± SD of five independent experiments are shown. * p
    Figure Legend Snippet: TLR2 engagement on CD4+ T cells enhances IL9 mRNA and protein expression driven by polyclonal activation and TGF-β and IL-4. Naïve CD4+T cells from WT mice were stimulated with anti-CD3 and anti-CD28 mAbs and exogenous TGF-β (5ng/ml) and IL-4 (10ng/ml), with or without P3CSK4 (1μg/ml) for 48h. (A) Relative expression of Il9 mRNA under polarizing conditions with or without P3CSK4 (n=3). (B) Flow cytometric analysis of CD4+ T cells labeled for intracellular IL-9 and IFN-γ. (C) IL-9 in culture supernatants was determined by ELISA. (D) IFN-γ in culture supernatants was determined by ELISA. Means ± SD of five independent experiments are shown. * p

    Techniques Used: Expressing, Activation Assay, Mouse Assay, Flow Cytometry, Labeling, Enzyme-linked Immunosorbent Assay

    TLR2 engagement on human CD4 + T cells enhances IL9 mRNA and protein expression driven by polyclonal activation and TGF-β and IL-4. Naïve CD4 + T cells from three human donors were stimulated with anti-CD3 (10ug/ml) and anti-CD28 mAbs (1ug/ml) and exogenous TGF-β (5ng/ml) and IL-4 (10ng/ml), with or without P3CSK4 (2μg/ml) for 48h. (A) Relative expression of Il9 mRNA under non-polarizing and polarizing conditions with or without P3CSK4 (n=3) is shown. (B) IL-9 cytokine in culture supernatants were determined by ELISA. Means ± SD of three technical replicates for each donor are shown.
    Figure Legend Snippet: TLR2 engagement on human CD4 + T cells enhances IL9 mRNA and protein expression driven by polyclonal activation and TGF-β and IL-4. Naïve CD4 + T cells from three human donors were stimulated with anti-CD3 (10ug/ml) and anti-CD28 mAbs (1ug/ml) and exogenous TGF-β (5ng/ml) and IL-4 (10ng/ml), with or without P3CSK4 (2μg/ml) for 48h. (A) Relative expression of Il9 mRNA under non-polarizing and polarizing conditions with or without P3CSK4 (n=3) is shown. (B) IL-9 cytokine in culture supernatants were determined by ELISA. Means ± SD of three technical replicates for each donor are shown.

    Techniques Used: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay

    10) Product Images from "Genetic polymorphism directs IL-6 expression in fibroblasts but not selected other cell types"

    Article Title: Genetic polymorphism directs IL-6 expression in fibroblasts but not selected other cell types

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1520861112

    Elevated unspliced mRNA levels in high-producing fibroblasts supports transcriptional regulation of the differential IL-6 response between synovial fibroblasts. Unspliced ( A and B ) and mature ( C ) IL-6 mRNA expression for two high IL-6–expressing (OA4, OA6) and two low-expressing (RA341, OA502) was determined by qRT-PCR and normalized to the housekeeping gene HPRT. Unspliced primer 1 ( A ) amplifies a region of intron 1. Unspliced primer 2 ( B ) amplifies a region of intron 2. Unspliced mRNA expression averaged 3.5 ± 1.6% of mature mRNA. Primers directed against the IL-6 proximal promoter showed low contamination with genomic DNA, averaging 1.7 ± 2.4% of unspliced mRNA.
    Figure Legend Snippet: Elevated unspliced mRNA levels in high-producing fibroblasts supports transcriptional regulation of the differential IL-6 response between synovial fibroblasts. Unspliced ( A and B ) and mature ( C ) IL-6 mRNA expression for two high IL-6–expressing (OA4, OA6) and two low-expressing (RA341, OA502) was determined by qRT-PCR and normalized to the housekeeping gene HPRT. Unspliced primer 1 ( A ) amplifies a region of intron 1. Unspliced primer 2 ( B ) amplifies a region of intron 2. Unspliced mRNA expression averaged 3.5 ± 1.6% of mature mRNA. Primers directed against the IL-6 proximal promoter showed low contamination with genomic DNA, averaging 1.7 ± 2.4% of unspliced mRNA.

    Techniques Used: Expressing, Quantitative RT-PCR

    11) Product Images from "Quantitative Analysis of Proteome Modulations in Alveolar Epithelial Type II Cells in Response to Pulmonary Aspergillus fumigatus Infection *"

    Article Title: Quantitative Analysis of Proteome Modulations in Alveolar Epithelial Type II Cells in Response to Pulmonary Aspergillus fumigatus Infection *

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1074/mcp.RA117.000072

    Main Protein Clusters Resulting from String Network Analysis. STRING network analysis was performed for significantly differentially abundant proteins ( n = 154) considering only highest confidence interactions (interaction score 0.9, text mining as a source of interaction was disabled). The analysis resulted in two main protein clusters, one of them representing proteins related to oxidative phosphorylation (blue circles). The second cluster (green circles) contained proteins related to oxidation-reduction processes, one of them the most strikingly upregulated protein IL4I1 (highlighted in red). Gene names are given next to the protein symbols, ratios of means are indicated next to the gene names, green arrows indicate higher protein abundance in the A. fumigatus infected group. Edges represent protein-protein interactions with the color code given next to the network.
    Figure Legend Snippet: Main Protein Clusters Resulting from String Network Analysis. STRING network analysis was performed for significantly differentially abundant proteins ( n = 154) considering only highest confidence interactions (interaction score 0.9, text mining as a source of interaction was disabled). The analysis resulted in two main protein clusters, one of them representing proteins related to oxidative phosphorylation (blue circles). The second cluster (green circles) contained proteins related to oxidation-reduction processes, one of them the most strikingly upregulated protein IL4I1 (highlighted in red). Gene names are given next to the protein symbols, ratios of means are indicated next to the gene names, green arrows indicate higher protein abundance in the A. fumigatus infected group. Edges represent protein-protein interactions with the color code given next to the network.

    Techniques Used: Infection

    12) Product Images from "Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties"

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0145749

    RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .
    Figure Legend Snippet: RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Agarose Gel Electrophoresis

    13) Product Images from "FITC Conjugation Markedly Enhances Hepatic Clearance of N-Formyl Peptides"

    Article Title: FITC Conjugation Markedly Enhances Hepatic Clearance of N-Formyl Peptides

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0160602

    Expression of FPR1 mRNA and protein in liver and isolated liver cells. (A) Total RNA was isolated from pelleted cells and whole tissue. cDNA was synthesized from 1 μg of total RNA using the QuantiTect Reverse Transcription Kit. PCR was performed in a 25 μl reaction mixture containing 2 μl of cDNA of hHeps (1), hLSECs (2), human liver (3), mHeps (4), mLSECs (5), and mouse liver (6). The house keeping genes were β-actin and GAPDH. (B) Homogenized tissue specimens and cells were lysed in a RIPA lysis buffer with protease and phosphatase inhibitor cocktails. Equal amounts of protein were loaded into NuPAGE Novex 4–12% Bis-Tris gel, then blotted onto PVDF membrane and probed with antibodies against FPR1 and β-actin.
    Figure Legend Snippet: Expression of FPR1 mRNA and protein in liver and isolated liver cells. (A) Total RNA was isolated from pelleted cells and whole tissue. cDNA was synthesized from 1 μg of total RNA using the QuantiTect Reverse Transcription Kit. PCR was performed in a 25 μl reaction mixture containing 2 μl of cDNA of hHeps (1), hLSECs (2), human liver (3), mHeps (4), mLSECs (5), and mouse liver (6). The house keeping genes were β-actin and GAPDH. (B) Homogenized tissue specimens and cells were lysed in a RIPA lysis buffer with protease and phosphatase inhibitor cocktails. Equal amounts of protein were loaded into NuPAGE Novex 4–12% Bis-Tris gel, then blotted onto PVDF membrane and probed with antibodies against FPR1 and β-actin.

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, Lysis

    14) Product Images from "Sox9 regulates cell state and activity of embryonic mouse mammary progenitor cells"

    Article Title: Sox9 regulates cell state and activity of embryonic mouse mammary progenitor cells

    Journal: Communications Biology

    doi: 10.1038/s42003-018-0215-3

    Effect of Sox9 ablation on embryonic mammary progenitor cell fate and function. a qRT-PCR for Sox9 , Zeb1, Snai2 , Twist , Procr , Trp63 , Acta2 , and Ecad in e1/Co#1 and e1/ Sox9 -KO#1 cells. Relative quantification of each sample is normalised to that of e1/Co#1 and shown in log 2 plot with error bar of triplicates ( n = 3, mean ± s.e.m.). b Heatmap from RNA-seq of e1/control and e1/ Sox9 -KO cells of genes that were significantly up- or down-regulated in all three e1/ Sox9 -KO subclones using both DESeq and Intensity Difference test. c In vitro alveologenesis assay results. Formation of alveoli-like structure and milk production was assessed by morphological changes using bright-field microscopy, as well as IF using anti-milk antibody (red) and DAPI. Scale bar, 200 μm. d In vitro alveologenesis assay results. Quantification of number of alveoli
    Figure Legend Snippet: Effect of Sox9 ablation on embryonic mammary progenitor cell fate and function. a qRT-PCR for Sox9 , Zeb1, Snai2 , Twist , Procr , Trp63 , Acta2 , and Ecad in e1/Co#1 and e1/ Sox9 -KO#1 cells. Relative quantification of each sample is normalised to that of e1/Co#1 and shown in log 2 plot with error bar of triplicates ( n = 3, mean ± s.e.m.). b Heatmap from RNA-seq of e1/control and e1/ Sox9 -KO cells of genes that were significantly up- or down-regulated in all three e1/ Sox9 -KO subclones using both DESeq and Intensity Difference test. c In vitro alveologenesis assay results. Formation of alveoli-like structure and milk production was assessed by morphological changes using bright-field microscopy, as well as IF using anti-milk antibody (red) and DAPI. Scale bar, 200 μm. d In vitro alveologenesis assay results. Quantification of number of alveoli

    Techniques Used: Quantitative RT-PCR, RNA Sequencing Assay, In Vitro, Microscopy

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    Amplification:

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    Synthesized:

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    Cytometry:

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    Quantitative RT-PCR:

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    SYBR Green Assay:

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    Incubation:

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    Expressing:

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    Gel Purification:

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    Flow Cytometry:

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    Sequencing:

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    Activation Assay:

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    Northern Blot:

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    Generated:

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    Reverse Transcription Polymerase Chain Reaction:

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    Hybridization:

    Article Title: HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ▿ ▿ †
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    Isolation:

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    Labeling:

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    Article Snippet: The labeling of the probe, hybridization of the blot, and detection of the signal were performed using the AlkPhos direct labeling and detection kit (GE Healthcare Life Sciences) according to the manufacturer's instructions. .. For synthesis of shorter cDNA fragments, the QuantiTect reverse transcription kit (Qiagen), which provides integrated genomic DNA removal, was used.

    Purification:

    Article Title: IL-38: A new factor in rheumatoid arthritis
    Article Snippet: .. For the follow-up real-time-quantitative PCR (qPCR) assays, purified RNA was converted in each instance into cDNA using a Quantitect reverse transcription kit (Qiagen). qPCRs were performed with SYBR Green Mastermix (Qiagen) using primers for mouse and human β-actin and IL-38 and mouse IL-1β and IL-6 (Qiagen) on an Mx3000p PCR machine (Stratagene, La Jolla, CA). .. Relative expression was calculated using the comparative threshold cycle method, as reported previously .

    Article Title: HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ▿ ▿ †
    Article Snippet: For RT-PCRs, 1 μg of purified total RNA was reverse transcribed with the MonsterScript reverse transcriptase kit, according to the manufacturer's protocol (Epicentre) using random primers. .. For synthesis of shorter cDNA fragments, the QuantiTect reverse transcription kit (Qiagen), which provides integrated genomic DNA removal, was used.

    Polymerase Chain Reaction:

    Article Title: HPV8 Field Cancerization in a Transgenic Mouse Model Is due to Lrig1+ Keratinocyte Stem Cell Expansion
    Article Snippet: RNA isolation from sorted cells and homogenized tissues was performed with an RNeasy kit and cDNA was synthetized using a QuantiTect Reverse Transcription Kit (Qiagen, Manchester, UK). .. For the determination of E6 and E7 gene expression levels, SyGreen (PCR Biosystems, London, UK) was used.

    Article Title: IL-38: A new factor in rheumatoid arthritis
    Article Snippet: .. For the follow-up real-time-quantitative PCR (qPCR) assays, purified RNA was converted in each instance into cDNA using a Quantitect reverse transcription kit (Qiagen). qPCRs were performed with SYBR Green Mastermix (Qiagen) using primers for mouse and human β-actin and IL-38 and mouse IL-1β and IL-6 (Qiagen) on an Mx3000p PCR machine (Stratagene, La Jolla, CA). .. Relative expression was calculated using the comparative threshold cycle method, as reported previously .

    Article Title: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
    Article Snippet: Complimentary DNA (cDNA) was generated (Qiagen QuantiTect Reverse Transcription Kit) and subjected to pre-amplification (SsoAdvanced PreAmp Supermix, Bio-Rad). .. The pre-amplified cDNA was subjected to qPCR (Qiagen SYBR Green PCR kit; 60 °C annealing temperature, 60 cycles).

    Article Title: Effects of Environmental Enrichment on Doublecortin and BDNF Expression along the Dorso-Ventral Axis of the Dentate Gyrus
    Article Snippet: Total RNA was reverse-transcribed using a QuantiTect reverse transcription kit (Qiagen) that incorporates a DNase treatment step. .. Standards were produced by gel purification of PCR products using a QiaQuick gel extraction kit (Qiagen) and their concentration was measured on a NanoDrop spectrophotometer (Thermo Fisher Scientific).

    FACS:

    Article Title: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
    Article Snippet: RNA was isolated from FACS-sorted CD31+ CD45− cells according to the manufacturer’s protocol (Qiagen mini-prep kit), and concentration and purity were determined using a NanoDrop Spectrophotometer (Thermo Fisher). .. Complimentary DNA (cDNA) was generated (Qiagen QuantiTect Reverse Transcription Kit) and subjected to pre-amplification (SsoAdvanced PreAmp Supermix, Bio-Rad).

    Gel Extraction:

    Article Title: Effects of Environmental Enrichment on Doublecortin and BDNF Expression along the Dorso-Ventral Axis of the Dentate Gyrus
    Article Snippet: Total RNA was reverse-transcribed using a QuantiTect reverse transcription kit (Qiagen) that incorporates a DNase treatment step. .. Standards were produced by gel purification of PCR products using a QiaQuick gel extraction kit (Qiagen) and their concentration was measured on a NanoDrop spectrophotometer (Thermo Fisher Scientific).

    Software:

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties
    Article Snippet: Expression studies by RT-PCR and RT-qPCR Expression studies of a few representative AFL and BRL genes relevant to the study’s objectives were performed by RT-PCR and RT-qPCR using the cDNA prepared with QuantiTect Reverse-Transcription Kit (Qiagen), as described above. .. Gene-specific primers not less than 20 nt long and with Tm values no less than 59.0°C were designed using the NCBI Primer Blast software ( http://www.ncbi.nlm.nih.gov/tools/primer-blast ).

    Real-time Polymerase Chain Reaction:

    Article Title: HPV8 Field Cancerization in a Transgenic Mouse Model Is due to Lrig1+ Keratinocyte Stem Cell Expansion
    Article Snippet: RNA isolation from sorted cells and homogenized tissues was performed with an RNeasy kit and cDNA was synthetized using a QuantiTect Reverse Transcription Kit (Qiagen, Manchester, UK). .. Real-time quantitative reverse transcription analysis was performed on the QuantStudio 7 Flex Real-Time PCR System (ThermoFisher Scientific).

    Article Title: IL-38: A new factor in rheumatoid arthritis
    Article Snippet: .. For the follow-up real-time-quantitative PCR (qPCR) assays, purified RNA was converted in each instance into cDNA using a Quantitect reverse transcription kit (Qiagen). qPCRs were performed with SYBR Green Mastermix (Qiagen) using primers for mouse and human β-actin and IL-38 and mouse IL-1β and IL-6 (Qiagen) on an Mx3000p PCR machine (Stratagene, La Jolla, CA). .. Relative expression was calculated using the comparative threshold cycle method, as reported previously .

    Article Title: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
    Article Snippet: Paragraph title: Quantitative PCR ... Complimentary DNA (cDNA) was generated (Qiagen QuantiTect Reverse Transcription Kit) and subjected to pre-amplification (SsoAdvanced PreAmp Supermix, Bio-Rad).

    Article Title: Effects of Environmental Enrichment on Doublecortin and BDNF Expression along the Dorso-Ventral Axis of the Dentate Gyrus
    Article Snippet: Total RNA was reverse-transcribed using a QuantiTect reverse transcription kit (Qiagen) that incorporates a DNase treatment step. .. We performed real-time PCR experiments using the Rotor-Disc® 100 on a Rotor-Gene® Q thermal cycler (Qiagen, Germany) using QuantiTect mouse primer mixes (Qiagen).

    Article Title: SOX7 Is Required for Muscle Satellite Cell Development and Maintenance
    Article Snippet: Paragraph title: Quantitative Real-Time PCR ... Micro-elute Total RNA Kit I as per the manufacturer's protocol (Omega Bio-tek) and reverse transcribed using a Quantitect Reverse Transcription Kit (QIAGEN).

    Article Title: Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival, et al. Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival
    Article Snippet: One micrograms of total RNA was reverse transcribed using the QuantiTect Reverse Transcription Kit (Qiagen). .. RTqPCR for IL‐1α, IL‐1β, IL‐2, IL‐6, IL‐10, IL‐17A, interferon γ (IFN‐γ), and tumor necrosis factor α (TNF‐α) was performed on a 7500 real‐time PCR system (Applied Biosystems) with Sybr green (Qiagen).

    Spectrophotometry:

    Article Title: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
    Article Snippet: RNA was isolated from FACS-sorted CD31+ CD45− cells according to the manufacturer’s protocol (Qiagen mini-prep kit), and concentration and purity were determined using a NanoDrop Spectrophotometer (Thermo Fisher). .. Complimentary DNA (cDNA) was generated (Qiagen QuantiTect Reverse Transcription Kit) and subjected to pre-amplification (SsoAdvanced PreAmp Supermix, Bio-Rad).

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties
    Article Snippet: The quality and quantity of the individual RNA preparations were determined using a NanoDrop spectrophotometer. .. QuantiTect Reverse Transcription Kit (Qiagen), which provides an optimized mixture of random primers and oligo-dT and gDNA Wipeout Buffer, was used to convert the RNA to cDNA.

    Article Title: FITC Conjugation Markedly Enhances Hepatic Clearance of N-Formyl Peptides
    Article Snippet: .. The quantity and quality of the extracted RNA was determined by the use of a spectrophotometer NanoDrop ND-1000 (Wilmington, DE, USA). cDNA was synthesized from 1 μg of total RNA using the QuantiTect Reverse Transcription Kit (Qiagen) according to the manufacturer's protocol. .. Quantitative PCR assay PCR was performed in a 25 μl reaction mixture containing 2 μl of cDNA (from isolated RNA), 12.5 μl of the JumpStart REDTaq® Ready Mix (Sigma-Aldrich), 0.5 μl of 10 μM forward and reverse primers, and 9.5 μl of ddH2 O.

    Article Title: Effects of Environmental Enrichment on Doublecortin and BDNF Expression along the Dorso-Ventral Axis of the Dentate Gyrus
    Article Snippet: Total RNA was reverse-transcribed using a QuantiTect reverse transcription kit (Qiagen) that incorporates a DNase treatment step. .. Standards were produced by gel purification of PCR products using a QiaQuick gel extraction kit (Qiagen) and their concentration was measured on a NanoDrop spectrophotometer (Thermo Fisher Scientific).

    Sampling:

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties
    Article Snippet: RNA isolation and cDNA preparation Total RNA from the superior and inferior spikelets collected on each sampling day and stored frozen at -80°C was extracted using TRIzol (Invitrogen, Life Technologies) following the manufacturer’s instructions. .. QuantiTect Reverse Transcription Kit (Qiagen), which provides an optimized mixture of random primers and oligo-dT and gDNA Wipeout Buffer, was used to convert the RNA to cDNA.

    Produced:

    Article Title: Effects of Environmental Enrichment on Doublecortin and BDNF Expression along the Dorso-Ventral Axis of the Dentate Gyrus
    Article Snippet: Total RNA was reverse-transcribed using a QuantiTect reverse transcription kit (Qiagen) that incorporates a DNase treatment step. .. Standards were produced by gel purification of PCR products using a QiaQuick gel extraction kit (Qiagen) and their concentration was measured on a NanoDrop spectrophotometer (Thermo Fisher Scientific).

    Concentration Assay:

    Article Title: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
    Article Snippet: RNA was isolated from FACS-sorted CD31+ CD45− cells according to the manufacturer’s protocol (Qiagen mini-prep kit), and concentration and purity were determined using a NanoDrop Spectrophotometer (Thermo Fisher). .. Complimentary DNA (cDNA) was generated (Qiagen QuantiTect Reverse Transcription Kit) and subjected to pre-amplification (SsoAdvanced PreAmp Supermix, Bio-Rad).

    Article Title: Effects of Environmental Enrichment on Doublecortin and BDNF Expression along the Dorso-Ventral Axis of the Dentate Gyrus
    Article Snippet: Total RNA was reverse-transcribed using a QuantiTect reverse transcription kit (Qiagen) that incorporates a DNase treatment step. .. Standards were produced by gel purification of PCR products using a QiaQuick gel extraction kit (Qiagen) and their concentration was measured on a NanoDrop spectrophotometer (Thermo Fisher Scientific).

    Lysis:

    Article Title: IL-38: A new factor in rheumatoid arthritis
    Article Snippet: The samples were treated with proteinase K (55 °C, 30 min), and cells were disrupted with Buffer RLT lysis buffer (Qiagen). .. For the follow-up real-time-quantitative PCR (qPCR) assays, purified RNA was converted in each instance into cDNA using a Quantitect reverse transcription kit (Qiagen). qPCRs were performed with SYBR Green Mastermix (Qiagen) using primers for mouse and human β-actin and IL-38 and mouse IL-1β and IL-6 (Qiagen) on an Mx3000p PCR machine (Stratagene, La Jolla, CA).

    Staining:

    Article Title: Identification of the S100 fused-type protein hornerin as a regulator of tumor vascularity
    Article Snippet: The cell suspension (filtrate) was incubated with the following antibody/dye panel for FACS; anti-CD45 AF488 (1:100) (Biolegend, San Diego, CA, USA, 103121, clone 30-F11), anti-CD31 AF647 (1:100) (Biolegend, 102415, clone 390), LIVE/DEAD violet stain (1:200) (Thermo Fisher). .. Complimentary DNA (cDNA) was generated (Qiagen QuantiTect Reverse Transcription Kit) and subjected to pre-amplification (SsoAdvanced PreAmp Supermix, Bio-Rad).

    T-Test:

    Article Title: Rapid reversal of innate immune dysregulation in blood of patients and livers of humanized mice with HCV following DAA therapy
    Article Snippet: A paired t-test was used to calculate p-values for all genes (22,283 transcripts) comparing pre- vs. post- interferon treatment. .. For RT-PCR, 1ug of mRNA was converted to cDNA using the QuantiTect Reverse Transcription kit (Qiagen) according to manufacturer protocol. cDNA was then analyzed for gene expression levels using the indicated primers (Qiagen)

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    Qiagen quantitect reverse transcription kit
    RT q PCR analysis showing relative gene expression levels of IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, TNF ‐α, and IFN ‐γ in allograft skin (A – H) and muscle (I – P). For RT q PCR , a total volume of 15 μl per reaction was used containing 50 ng cDNA template, 1.5 μl <t>QuantiTect</t> Primer Assay (Qiagen), 7.5 μl QuantiTect SYBR green PCR kit (Qiagen), and 5 μl ddH2O. Primers for IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, IFN ‐γ, and TNF ‐α were obtained from Qiagen. Cycling conditions included a hot start activation (5 minutes, 95°C), 40 cycles of 10 seconds denaturation (95°C), and annealing and extension (30 seconds, 60°C). Amplicons were quantified with the comparative threshold cycle ( CT ) method; data acquisition was performed using the 7500 System SDS Software Version 2.0.5 (Applied Biosystems). Amplification specificity was checked using melting curve according to the manufacturer's instructions. All samples were measured in duplicate and respective results were averaged. A–P, In allograft skin (A–H) and muscle (I–P) no significant differences in relative gene expression of cytokines was observed between anti‐ IL ‐1β‐treatment groups 3 and 4 and group 2 controls. Expression levels of all markers were normalized to those of group 2
    Quantitect Reverse Transcription Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2605 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    RT q PCR analysis showing relative gene expression levels of IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, TNF ‐α, and IFN ‐γ in allograft skin (A – H) and muscle (I – P). For RT q PCR , a total volume of 15 μl per reaction was used containing 50 ng cDNA template, 1.5 μl QuantiTect Primer Assay (Qiagen), 7.5 μl QuantiTect SYBR green PCR kit (Qiagen), and 5 μl ddH2O. Primers for IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, IFN ‐γ, and TNF ‐α were obtained from Qiagen. Cycling conditions included a hot start activation (5 minutes, 95°C), 40 cycles of 10 seconds denaturation (95°C), and annealing and extension (30 seconds, 60°C). Amplicons were quantified with the comparative threshold cycle ( CT ) method; data acquisition was performed using the 7500 System SDS Software Version 2.0.5 (Applied Biosystems). Amplification specificity was checked using melting curve according to the manufacturer's instructions. All samples were measured in duplicate and respective results were averaged. A–P, In allograft skin (A–H) and muscle (I–P) no significant differences in relative gene expression of cytokines was observed between anti‐ IL ‐1β‐treatment groups 3 and 4 and group 2 controls. Expression levels of all markers were normalized to those of group 2

    Journal: American Journal of Transplantation

    Article Title: Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival, et al. Subcutaneous administration of a neutralizing IL‐1β antibody prolongs limb allograft survival

    doi: 10.1111/ajt.14765

    Figure Lengend Snippet: RT q PCR analysis showing relative gene expression levels of IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, TNF ‐α, and IFN ‐γ in allograft skin (A – H) and muscle (I – P). For RT q PCR , a total volume of 15 μl per reaction was used containing 50 ng cDNA template, 1.5 μl QuantiTect Primer Assay (Qiagen), 7.5 μl QuantiTect SYBR green PCR kit (Qiagen), and 5 μl ddH2O. Primers for IL ‐1α, IL ‐1β, IL ‐2, IL ‐6, IL ‐10, IL ‐17A, IFN ‐γ, and TNF ‐α were obtained from Qiagen. Cycling conditions included a hot start activation (5 minutes, 95°C), 40 cycles of 10 seconds denaturation (95°C), and annealing and extension (30 seconds, 60°C). Amplicons were quantified with the comparative threshold cycle ( CT ) method; data acquisition was performed using the 7500 System SDS Software Version 2.0.5 (Applied Biosystems). Amplification specificity was checked using melting curve according to the manufacturer's instructions. All samples were measured in duplicate and respective results were averaged. A–P, In allograft skin (A–H) and muscle (I–P) no significant differences in relative gene expression of cytokines was observed between anti‐ IL ‐1β‐treatment groups 3 and 4 and group 2 controls. Expression levels of all markers were normalized to those of group 2

    Article Snippet: One micrograms of total RNA was reverse transcribed using the QuantiTect Reverse Transcription Kit (Qiagen).

    Techniques: Polymerase Chain Reaction, Expressing, SYBR Green Assay, Activation Assay, Software, Amplification

    RT-PCR analysis to evaluate transcriptional linkage of PG0104 to the 5′ end of the capsule operon and transcription through the hairpin region. Total RNA was isolated from the wild-type strain, and cDNA was generated as described in Materials and Methods with the QuantiTect reverse transcription kit. In the subsequent PCRs, the following primer combinations were used: RT_end104 (lane 1), RT_link104 (lane 2), RT_hairpin (lane 3), RT_link106 (lane 4), RT_start106 (lane 5). The figure demonstrates linkage of PG0104 to the hairpin, transcription of the hairpin, and linkage of the hairpin to PG0106. M, DNA size markers.

    Journal: Journal of Bacteriology

    Article Title: HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ▿ ▿ †

    doi: 10.1128/JB.00106-10

    Figure Lengend Snippet: RT-PCR analysis to evaluate transcriptional linkage of PG0104 to the 5′ end of the capsule operon and transcription through the hairpin region. Total RNA was isolated from the wild-type strain, and cDNA was generated as described in Materials and Methods with the QuantiTect reverse transcription kit. In the subsequent PCRs, the following primer combinations were used: RT_end104 (lane 1), RT_link104 (lane 2), RT_hairpin (lane 3), RT_link106 (lane 4), RT_start106 (lane 5). The figure demonstrates linkage of PG0104 to the hairpin, transcription of the hairpin, and linkage of the hairpin to PG0106. M, DNA size markers.

    Article Snippet: For synthesis of shorter cDNA fragments, the QuantiTect reverse transcription kit (Qiagen), which provides integrated genomic DNA removal, was used.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Generated

    RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .

    Journal: PLoS ONE

    Article Title: Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties

    doi: 10.1371/journal.pone.0145749

    Figure Lengend Snippet: RT-PCR of total RNA isolated from superior (A) and inferior (B) spikelets of O. sativa cv. Mahalaxmi panicles on 0, 3 and 6 days after anthesis (DAA or simply D) for analysis of genes overexpressed in the superior compared with inferior spikelets (I) and genes overexpressed in the inferior compared with superior spikelets (II). Total RNA was isolated using TRIzol (Life Technologies) and reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen). Each gene was amplified using gene-specific primers designed using Primer Blast. Actin and 18S rRNA were amplified for 30 and 25 cycles, respectively, as positive and loading controls. The other reactions were performed for 30 cycles, and the amplified products were separated on an agarose gel containing ETBR and visualized and photographed using a Gel Doc (Bio-Rad). The primer sequences are provided in S2 Table .

    Article Snippet: QuantiTect Reverse Transcription Kit (Qiagen), which provides an optimized mixture of random primers and oligo-dT and gDNA Wipeout Buffer, was used to convert the RNA to cDNA.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Agarose Gel Electrophoresis

    Elevated unspliced mRNA levels in high-producing fibroblasts supports transcriptional regulation of the differential IL-6 response between synovial fibroblasts. Unspliced ( A and B ) and mature ( C ) IL-6 mRNA expression for two high IL-6–expressing (OA4, OA6) and two low-expressing (RA341, OA502) was determined by qRT-PCR and normalized to the housekeeping gene HPRT. Unspliced primer 1 ( A ) amplifies a region of intron 1. Unspliced primer 2 ( B ) amplifies a region of intron 2. Unspliced mRNA expression averaged 3.5 ± 1.6% of mature mRNA. Primers directed against the IL-6 proximal promoter showed low contamination with genomic DNA, averaging 1.7 ± 2.4% of unspliced mRNA.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Genetic polymorphism directs IL-6 expression in fibroblasts but not selected other cell types

    doi: 10.1073/pnas.1520861112

    Figure Lengend Snippet: Elevated unspliced mRNA levels in high-producing fibroblasts supports transcriptional regulation of the differential IL-6 response between synovial fibroblasts. Unspliced ( A and B ) and mature ( C ) IL-6 mRNA expression for two high IL-6–expressing (OA4, OA6) and two low-expressing (RA341, OA502) was determined by qRT-PCR and normalized to the housekeeping gene HPRT. Unspliced primer 1 ( A ) amplifies a region of intron 1. Unspliced primer 2 ( B ) amplifies a region of intron 2. Unspliced mRNA expression averaged 3.5 ± 1.6% of mature mRNA. Primers directed against the IL-6 proximal promoter showed low contamination with genomic DNA, averaging 1.7 ± 2.4% of unspliced mRNA.

    Article Snippet: RNA was isolated by using manufacturer’s instructions (RNeasy Micro kit; Qiagen) and analyzed by quantitative reverse-transcription PCR (qRT-PCR; QuantiTect Reverse Transcription kit, Qiagen; Brilliant III Ultra-Fast SYBR Green, Agilent).

    Techniques: Expressing, Quantitative RT-PCR