quantitect taqman probe rt pcr kit  (Qiagen)

 
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    Name:
    QuantiTect Probe RT PCR Kit
    Description:
    For one step qRT PCR using sequence specific probes for gene expression analysis Kit contents Qiagen QuantiTect Probe RT PCR Kit 200 x 50L rxns RNA Targets Sample One step RT PCR Reaction Sequence specific Probes Ideal for Real time Quantification of RNA Targets For One step qRT PCR Using Sequence specific Probes for Gene Expression Analysis Includes 3 x 1 7mL 2x Qiagen QuantiTect Probe RT PCR Master Mix 100L Qiagen QuantiTect RT Mix 2 x 2mL RNase free Water Benefits Highly sensitive detection of low copy targets Accurate quantification over several logs of template Use of any sequence specific probe on any real time cycler No need to optimize reaction and cycling conditions
    Catalog Number:
    204443
    Price:
    645
    Category:
    QuantiTect Probe RT PCR Kit
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    Structured Review

    Qiagen quantitect taqman probe rt pcr kit
    QuantiTect Probe RT PCR Kit
    For one step qRT PCR using sequence specific probes for gene expression analysis Kit contents Qiagen QuantiTect Probe RT PCR Kit 200 x 50L rxns RNA Targets Sample One step RT PCR Reaction Sequence specific Probes Ideal for Real time Quantification of RNA Targets For One step qRT PCR Using Sequence specific Probes for Gene Expression Analysis Includes 3 x 1 7mL 2x Qiagen QuantiTect Probe RT PCR Master Mix 100L Qiagen QuantiTect RT Mix 2 x 2mL RNase free Water Benefits Highly sensitive detection of low copy targets Accurate quantification over several logs of template Use of any sequence specific probe on any real time cycler No need to optimize reaction and cycling conditions
    https://www.bioz.com/result/quantitect taqman probe rt pcr kit/product/Qiagen
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    Images

    1) Product Images from "Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis"

    Article Title: Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis

    Journal:

    doi: 10.1099/mic.0.2008/022293-0

    Quantitative TaqMan RT-PCR
    Figure Legend Snippet: Quantitative TaqMan RT-PCR

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    2) Product Images from "A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses"

    Article Title: A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses

    Journal: Journal of Medical Virology

    doi: 10.1002/jmv.21962

    Establishment and validation of multivirus real‐time PCR. A : Procedure of the multivirus real‐time PCR system. DNA sample was mixed with Quantitect 2× master mix, and RNA sample was mixed with Quantitect 2× master mix and reverse transcriptase (RT) mix. These mixtures were poured into each well in a 96‐well plate at 10 µl per well. Ten microliters of 2× probe and primer mix were then added to each well in a premixed 96‐well plate. Finally, the virus genes were amplified and detected in a real‐time PCR machine for 2 hr. B : Validation of the multivirus real‐time PCR. A gene expression image by TreeView software based on the results of the multivirus real‐time PCR for control samples is shown. A horizontal line shows each probe–primer set and a vertical line is one sample of positive control. Gray vertical lines indicate no sample. A scale bar indicates copy number of color. A green box indicates specific reactions of target positive controls in specific probe–primer sets. Arrows of (a–c) also show specific signals. The arrow (a) shows positive signal for TTV in a brain sample with both JCV and TTV infection. The arrows (b1–3) show that a probe–primer set for pan‐enterovirus reacted with poliovirus (b1), Coxsackievirus B3 (b2), and Echovirus 6 (b3) positive samples. The arrow (c) shows that a probe–primer set for influenza virus A reacted with H5N1 influenza virus. Details of positive controls were listed in Supplementary Table II.
    Figure Legend Snippet: Establishment and validation of multivirus real‐time PCR. A : Procedure of the multivirus real‐time PCR system. DNA sample was mixed with Quantitect 2× master mix, and RNA sample was mixed with Quantitect 2× master mix and reverse transcriptase (RT) mix. These mixtures were poured into each well in a 96‐well plate at 10 µl per well. Ten microliters of 2× probe and primer mix were then added to each well in a premixed 96‐well plate. Finally, the virus genes were amplified and detected in a real‐time PCR machine for 2 hr. B : Validation of the multivirus real‐time PCR. A gene expression image by TreeView software based on the results of the multivirus real‐time PCR for control samples is shown. A horizontal line shows each probe–primer set and a vertical line is one sample of positive control. Gray vertical lines indicate no sample. A scale bar indicates copy number of color. A green box indicates specific reactions of target positive controls in specific probe–primer sets. Arrows of (a–c) also show specific signals. The arrow (a) shows positive signal for TTV in a brain sample with both JCV and TTV infection. The arrows (b1–3) show that a probe–primer set for pan‐enterovirus reacted with poliovirus (b1), Coxsackievirus B3 (b2), and Echovirus 6 (b3) positive samples. The arrow (c) shows that a probe–primer set for influenza virus A reacted with H5N1 influenza virus. Details of positive controls were listed in Supplementary Table II.

    Techniques Used: Real-time Polymerase Chain Reaction, Amplification, Expressing, Software, Positive Control, Infection

    3) Product Images from "Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus"

    Article Title: Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus

    Journal: Journal of food protection

    doi: 10.4315/0362-028X.JFP-12-361

    qRT-PCR
    Figure Legend Snippet: qRT-PCR

    Techniques Used: Quantitative RT-PCR

    4) Product Images from "Chronic stress enhances progression of acute lymphoblastic leukemia via ?-adrenergic signaling"

    Article Title: Chronic stress enhances progression of acute lymphoblastic leukemia via ?-adrenergic signaling

    Journal: Brain, Behavior, and Immunity

    doi: 10.1016/j.bbi.2012.01.013

    Effect of chronic restraint stress on ALL progression. (A) 21-day longitudinal growth curves for total body Nalm-6 tumor burden in control and restraint-stressed animals. (B) Nalm-6 tumor burden in separate body areas at Day 14 in control and restraint-stressed
    Figure Legend Snippet: Effect of chronic restraint stress on ALL progression. (A) 21-day longitudinal growth curves for total body Nalm-6 tumor burden in control and restraint-stressed animals. (B) Nalm-6 tumor burden in separate body areas at Day 14 in control and restraint-stressed

    Techniques Used:

    Nalm-6 ALL Model. (A) Nalm-6 pre-B ALL cells were localized and quantified by periodic imaging of leukemia-specific bioluminescence signal from ventral and dorsal sides of SCID mice. (B) Femoral bone marrow white blood cells (WBC) and CD10+ Nalm-6 ALL
    Figure Legend Snippet: Nalm-6 ALL Model. (A) Nalm-6 pre-B ALL cells were localized and quantified by periodic imaging of leukemia-specific bioluminescence signal from ventral and dorsal sides of SCID mice. (B) Femoral bone marrow white blood cells (WBC) and CD10+ Nalm-6 ALL

    Techniques Used: Imaging, Mouse Assay

    β-adrenergic signaling in stress-enhanced ALL progression. (A) Nalm-6 ALL was quantified in control mice vs. stress mice that were treated with or without the β-blocker, propranolol; data representative of 3 independent experiments. (B)
    Figure Legend Snippet: β-adrenergic signaling in stress-enhanced ALL progression. (A) Nalm-6 ALL was quantified in control mice vs. stress mice that were treated with or without the β-blocker, propranolol; data representative of 3 independent experiments. (B)

    Techniques Used: Mouse Assay

    5) Product Images from "Enhanced pathogenicity of low-pathogenic H9N2 avian influenza virus after vaccination with infectious bronchitis live attenuated vaccine"

    Article Title: Enhanced pathogenicity of low-pathogenic H9N2 avian influenza virus after vaccination with infectious bronchitis live attenuated vaccine

    Journal: Veterinary World

    doi: 10.14202/vetworld.2018.977-985

    Column chart showing RNA copies of H9 using rRT-polymerase chain reaction in chicken groups and different time conditions. (a) Experiment 1, (b) Experiment 2.
    Figure Legend Snippet: Column chart showing RNA copies of H9 using rRT-polymerase chain reaction in chicken groups and different time conditions. (a) Experiment 1, (b) Experiment 2.

    Techniques Used: Polymerase Chain Reaction

    6) Product Images from "β-adrenergic-stimulated macrophages: Comprehensive localization in the M1–M2 spectrum"

    Article Title: β-adrenergic-stimulated macrophages: Comprehensive localization in the M1–M2 spectrum

    Journal: Brain, behavior, and immunity

    doi: 10.1016/j.bbi.2016.07.162

    Promoter-based bioinformatic analysis of transcription factors in genes differentially-regulated in β-adrenergic-stimulated vs. control macrophages. Data represent mean fold difference (mean ratio of isoproterenol/control) ±SE of transcription
    Figure Legend Snippet: Promoter-based bioinformatic analysis of transcription factors in genes differentially-regulated in β-adrenergic-stimulated vs. control macrophages. Data represent mean fold difference (mean ratio of isoproterenol/control) ±SE of transcription

    Techniques Used:

    Effect of β-adrenergic signaling on select gene transcripts that constitute macrophage activation categories along the M1–M2 spectrum in Murray et al., 2014. Isoproterenol at 1 μM. Data represent mean ± SE of three independent
    Figure Legend Snippet: Effect of β-adrenergic signaling on select gene transcripts that constitute macrophage activation categories along the M1–M2 spectrum in Murray et al., 2014. Isoproterenol at 1 μM. Data represent mean ± SE of three independent

    Techniques Used: Activation Assay

    Mean diagnosticity score for genes that were up-regulated or down-regulated by β-adrenergic signaling. Diagnosticity scores for each gene transcript quantified the extent to which that transcript was predominately expressed by an M2-polarized
    Figure Legend Snippet: Mean diagnosticity score for genes that were up-regulated or down-regulated by β-adrenergic signaling. Diagnosticity scores for each gene transcript quantified the extent to which that transcript was predominately expressed by an M2-polarized

    Techniques Used:

    7) Product Images from "β-adrenergic-stimulated macrophages: Comprehensive localization in the M1–M2 spectrum"

    Article Title: β-adrenergic-stimulated macrophages: Comprehensive localization in the M1–M2 spectrum

    Journal: Brain, behavior, and immunity

    doi: 10.1016/j.bbi.2016.07.162

    Effect of β-adrenergic signaling on select gene transcripts that constitute macrophage activation categories along the M1–M2 spectrum in Murray et al., 2014. Isoproterenol at 1 μM. Data represent mean ± SE of three independent
    Figure Legend Snippet: Effect of β-adrenergic signaling on select gene transcripts that constitute macrophage activation categories along the M1–M2 spectrum in Murray et al., 2014. Isoproterenol at 1 μM. Data represent mean ± SE of three independent

    Techniques Used: Activation Assay

    (A) M1 - M2 spectrum categories of macrophage activation, based on the activator and subsequent known transcription factors, as proposed by Peter J. Murray and colleagues following the International Congress of Immunology in Milan in 2013. At the far
    Figure Legend Snippet: (A) M1 - M2 spectrum categories of macrophage activation, based on the activator and subsequent known transcription factors, as proposed by Peter J. Murray and colleagues following the International Congress of Immunology in Milan in 2013. At the far

    Techniques Used: Activation Assay

    8) Product Images from "Integrated exome and transcriptome sequencing reveals ZAK isoform usage in gastric cancer"

    Article Title: Integrated exome and transcriptome sequencing reveals ZAK isoform usage in gastric cancer

    Journal: Nature Communications

    doi: 10.1038/ncomms4830

    Differential ZAK isoform usage between normal and tumour samples. ( a ) ZAK gene model and protein domain structure. Thick bars: coding exons; thin bars: UTR. TV2 lacks the last nine exons and has a long terminal coding and non-coding exon. Blue and red indicate unique TV1 and TV2 sequences. Transcript variant 1 (TV1) encodes a longer protein product with a sterile alpha motif domain. ( b ) ZAK TV1 fraction is significantly higher in gastric tumours (left) and colon tumours (right), compared with normal adjacent tissues. The fraction of TV1 was measured by the ratio between the number of reads uniquely assignable to TV1 and the number of reads mapped to the entire ZAK gene. To account for smooth muscle contamination in normal tissues, we fit a linear model with smoothelin expression as predictor and the log isoform fraction as response, and used the residuals of the model as the ‘adjusted isoform fraction’. Dots represent samples. Grey lines connect matched tumour and normal samples. The boxes in the box-and-whisker plots represent the interquartile range between the first and third quartiles; the dashed lines (whiskers) extend to the most extreme data points, which is no more than 1.5 times the interquartile range from the box. ( c ) ZAK isoform fractions derived from RNAseq data correlate with quantitative PCR (qPCR) measurements. For nine gastric cancer cell lines in our study, we quantified the ratio between ZAK total expression and ZAK TV1 expression using qPCR, and compared the measurements with the isoform fraction we derived from the RNAseq data. The two measurements have significant correlation (Pearson’s correlation coefficient r =0.91, P -value=0.00077). ( d ) ZAK isoform expression in six TCGA data sets where there are > 10 normal samples. Normal samples are represented by blue dots and tumour samples by red dots. ZAK TV1 fraction is significantly higher (adjusted P -value
    Figure Legend Snippet: Differential ZAK isoform usage between normal and tumour samples. ( a ) ZAK gene model and protein domain structure. Thick bars: coding exons; thin bars: UTR. TV2 lacks the last nine exons and has a long terminal coding and non-coding exon. Blue and red indicate unique TV1 and TV2 sequences. Transcript variant 1 (TV1) encodes a longer protein product with a sterile alpha motif domain. ( b ) ZAK TV1 fraction is significantly higher in gastric tumours (left) and colon tumours (right), compared with normal adjacent tissues. The fraction of TV1 was measured by the ratio between the number of reads uniquely assignable to TV1 and the number of reads mapped to the entire ZAK gene. To account for smooth muscle contamination in normal tissues, we fit a linear model with smoothelin expression as predictor and the log isoform fraction as response, and used the residuals of the model as the ‘adjusted isoform fraction’. Dots represent samples. Grey lines connect matched tumour and normal samples. The boxes in the box-and-whisker plots represent the interquartile range between the first and third quartiles; the dashed lines (whiskers) extend to the most extreme data points, which is no more than 1.5 times the interquartile range from the box. ( c ) ZAK isoform fractions derived from RNAseq data correlate with quantitative PCR (qPCR) measurements. For nine gastric cancer cell lines in our study, we quantified the ratio between ZAK total expression and ZAK TV1 expression using qPCR, and compared the measurements with the isoform fraction we derived from the RNAseq data. The two measurements have significant correlation (Pearson’s correlation coefficient r =0.91, P -value=0.00077). ( d ) ZAK isoform expression in six TCGA data sets where there are > 10 normal samples. Normal samples are represented by blue dots and tumour samples by red dots. ZAK TV1 fraction is significantly higher (adjusted P -value

    Techniques Used: Variant Assay, Expressing, Whisker Assay, Derivative Assay, Real-time Polymerase Chain Reaction

    Experimental validation of ZAK function in cancer. ( a ) Immunoblots of ZAK TV1 and TV2 expressions show that protein level of TV1 is higher in gastric tumours and cell lines, compared with normal stomach tissues. ZAK TV1 was detected with Bethyl α-ZAK antibody and TV2 with Sigma α-ZAK antibody (see Methods). ( b ) ZAK TV1, but not TV2, can stimulate multiple transcriptional programs related to cancer pathways. Transcription reporter assay in 293 cells transfected with empty vector, TV1 or TV2 along with the indicated firefly luciferase reporter construct (AP1, NFkB and TCF/LEF). Activity is normalized to cell number using CellTiter-Glo. Immunoblot shows relative ZAK isoform expression from 293 cells transfected with the indicated construct. ZAK was detected with Sigma α-ZAK antibody. ( c ) Depletion of ZAK from gastric cancer cell lines inhibits cell growth. In cell lines where ZAK knockdown led to reduced viability, there was consistently high TV1 expression, while TV2 expression was marginal and variable (for example, IM-95m cell line, see for example, panel a ). Cell viability analysis was carried out 6 days after infection of gastric cancer cell lines with independent ZAK shRNAs. Cell number is normalized to shNTC-infected cells. Immunoblot indicates the level of ZAK-TV1 depletion 4 days after infection. ZAK was detected with Bethyl α-ZAK antibody.
    Figure Legend Snippet: Experimental validation of ZAK function in cancer. ( a ) Immunoblots of ZAK TV1 and TV2 expressions show that protein level of TV1 is higher in gastric tumours and cell lines, compared with normal stomach tissues. ZAK TV1 was detected with Bethyl α-ZAK antibody and TV2 with Sigma α-ZAK antibody (see Methods). ( b ) ZAK TV1, but not TV2, can stimulate multiple transcriptional programs related to cancer pathways. Transcription reporter assay in 293 cells transfected with empty vector, TV1 or TV2 along with the indicated firefly luciferase reporter construct (AP1, NFkB and TCF/LEF). Activity is normalized to cell number using CellTiter-Glo. Immunoblot shows relative ZAK isoform expression from 293 cells transfected with the indicated construct. ZAK was detected with Sigma α-ZAK antibody. ( c ) Depletion of ZAK from gastric cancer cell lines inhibits cell growth. In cell lines where ZAK knockdown led to reduced viability, there was consistently high TV1 expression, while TV2 expression was marginal and variable (for example, IM-95m cell line, see for example, panel a ). Cell viability analysis was carried out 6 days after infection of gastric cancer cell lines with independent ZAK shRNAs. Cell number is normalized to shNTC-infected cells. Immunoblot indicates the level of ZAK-TV1 depletion 4 days after infection. ZAK was detected with Bethyl α-ZAK antibody.

    Techniques Used: Western Blot, Reporter Assay, Transfection, Plasmid Preparation, Luciferase, Construct, Activity Assay, Expressing, Infection

    9) Product Images from "The Open Reading Frame 3a Protein of Severe Acute Respiratory Syndrome-Associated Coronavirus Promotes Membrane Rearrangement and Cell Death ▿The Open Reading Frame 3a Protein of Severe Acute Respiratory Syndrome-Associated Coronavirus Promotes Membrane Rearrangement and Cell Death ▿ ‡"

    Article Title: The Open Reading Frame 3a Protein of Severe Acute Respiratory Syndrome-Associated Coronavirus Promotes Membrane Rearrangement and Cell Death ▿The Open Reading Frame 3a Protein of Severe Acute Respiratory Syndrome-Associated Coronavirus Promotes Membrane Rearrangement and Cell Death ▿ ‡

    Journal: Journal of Virology

    doi: 10.1128/JVI.01662-09

    SARS-CoV 3a-GFP partially colocalizes with the Golgi marker Gal-CFP. (A) Vero cells were cotransfected with 3a-GFP and Gal-CFP and analyzed at 24 hpt by confocal microscopy. 3a-GFP (A) and Gal-CFP (B) can be seen to partially colocalize in the merged
    Figure Legend Snippet: SARS-CoV 3a-GFP partially colocalizes with the Golgi marker Gal-CFP. (A) Vero cells were cotransfected with 3a-GFP and Gal-CFP and analyzed at 24 hpt by confocal microscopy. 3a-GFP (A) and Gal-CFP (B) can be seen to partially colocalize in the merged

    Techniques Used: Marker, Confocal Microscopy

    Expression of SARS-CoV 3a is sufficient to cause cell death, vesicle formation, and Golgi fragmentation. (A) Vero cells transfected with 3a-GFP were examined by confocal microscopy 48 hpt after propidium iodide (PI) staining to identify dead cells. (B)
    Figure Legend Snippet: Expression of SARS-CoV 3a is sufficient to cause cell death, vesicle formation, and Golgi fragmentation. (A) Vero cells transfected with 3a-GFP were examined by confocal microscopy 48 hpt after propidium iodide (PI) staining to identify dead cells. (B)

    Techniques Used: Expressing, Transfection, Confocal Microscopy, Staining

    SARS-CoV-infected cells exhibit cytoplamsic vesicles. (A and B) Vero cells were infected for 48 h with WT SARS-CoV and were fixed and analyzed by TEM. Infected cells show numerous vesicles and vacuoles and do not show characteristics of apoptosis such
    Figure Legend Snippet: SARS-CoV-infected cells exhibit cytoplamsic vesicles. (A and B) Vero cells were infected for 48 h with WT SARS-CoV and were fixed and analyzed by TEM. Infected cells show numerous vesicles and vacuoles and do not show characteristics of apoptosis such

    Techniques Used: Infection, Transmission Electron Microscopy

    Cell death caused by SARS-CoV is reduced by deletion of 3a. (A) Vero cells were mock infected (MOCK) or infected with wild-type (WT) or 3a-deficient (Δ3a) SARS-CoV. After 48 h, cells were examined by phase-contrast light microscopy. (B) Quantification
    Figure Legend Snippet: Cell death caused by SARS-CoV is reduced by deletion of 3a. (A) Vero cells were mock infected (MOCK) or infected with wild-type (WT) or 3a-deficient (Δ3a) SARS-CoV. After 48 h, cells were examined by phase-contrast light microscopy. (B) Quantification

    Techniques Used: Infection, Light Microscopy

    Membrane rearrangement and vesicle formation by SARS-CoV is dependent on 3a. (A) Vero cells were infected with WT SARS-CoV and 24 hpi were fixed and analyzed by TEM. Boxed region contains paranuclear vesicles. N, nucleus. (B) TEM of Vero cells either
    Figure Legend Snippet: Membrane rearrangement and vesicle formation by SARS-CoV is dependent on 3a. (A) Vero cells were infected with WT SARS-CoV and 24 hpi were fixed and analyzed by TEM. Boxed region contains paranuclear vesicles. N, nucleus. (B) TEM of Vero cells either

    Techniques Used: Infection, Transmission Electron Microscopy

    Golgi fragmentation caused by SARS-CoV infection depends on 3a. (A) Vero cells were transfected with Golgi marker Gal-CFP and after 24 h were mock infected (MOCK) or infected with SARS-CoV (WT) or Δ3a SARS-CoV (Δ3a) at a multiplicity of
    Figure Legend Snippet: Golgi fragmentation caused by SARS-CoV infection depends on 3a. (A) Vero cells were transfected with Golgi marker Gal-CFP and after 24 h were mock infected (MOCK) or infected with SARS-CoV (WT) or Δ3a SARS-CoV (Δ3a) at a multiplicity of

    Techniques Used: Infection, Transfection, Marker

    10) Product Images from "Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis"

    Article Title: Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-9-122

    Pharmacodynamic effects of TGF-β antagonists in vivo . A . Protein extracts from snap-frozen uninvolved lung tissue of mice treated with vehicle and LY2109761 and of mice treated with controls and 1D11 were prepared and subject to Western blotting using rabbit phosphoSmad2 antibody (1:1000 dilution). Whole cell lysate from SCP2TR was used as a control (Co). Treatment with both antagonists resulted in a reduction of phosphorylated Smad2 levels. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. B . RNA was extracted from snap-frozen uninvolved kidneys of mice treated with vehicle or LY2109761 using Trizol reagent (Invitrogen) and purified using RNeasy mini columns (Qiagen) according to the manufacturer's instructions. Transcript levels of CTGF and PAI-1 were assayed using the QuantiTect™ Probe RT-PCR Kit on a Mx4000 ® Multiplex Quantitative PCR System (Stratagene). Treatment with LY2109761 significantly reduced mRNA levels of both CTGF and PAI-1 relative to GAPDH mRNA. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. C . RNA was extracted from snap-frozen lungs of mice treated with vehicle, isotype control antibody or 1D11 and transcript levels determined as described above. No significant reduction in CTGF or PAI-1 mRNA levels could be detected in the 1D11-treated group. Values represent the means and SD of two independent experiments, 3 mice per group. Unpaired 2-sided t-test was used for comparisons.
    Figure Legend Snippet: Pharmacodynamic effects of TGF-β antagonists in vivo . A . Protein extracts from snap-frozen uninvolved lung tissue of mice treated with vehicle and LY2109761 and of mice treated with controls and 1D11 were prepared and subject to Western blotting using rabbit phosphoSmad2 antibody (1:1000 dilution). Whole cell lysate from SCP2TR was used as a control (Co). Treatment with both antagonists resulted in a reduction of phosphorylated Smad2 levels. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. B . RNA was extracted from snap-frozen uninvolved kidneys of mice treated with vehicle or LY2109761 using Trizol reagent (Invitrogen) and purified using RNeasy mini columns (Qiagen) according to the manufacturer's instructions. Transcript levels of CTGF and PAI-1 were assayed using the QuantiTect™ Probe RT-PCR Kit on a Mx4000 ® Multiplex Quantitative PCR System (Stratagene). Treatment with LY2109761 significantly reduced mRNA levels of both CTGF and PAI-1 relative to GAPDH mRNA. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. C . RNA was extracted from snap-frozen lungs of mice treated with vehicle, isotype control antibody or 1D11 and transcript levels determined as described above. No significant reduction in CTGF or PAI-1 mRNA levels could be detected in the 1D11-treated group. Values represent the means and SD of two independent experiments, 3 mice per group. Unpaired 2-sided t-test was used for comparisons.

    Techniques Used: In Vivo, Mouse Assay, Western Blot, Purification, Reverse Transcription Polymerase Chain Reaction, Multiplex Assay, Real-time Polymerase Chain Reaction

    11) Product Images from "Characterization of H5N1 Influenza Virus Variants with Hemagglutinin Mutations Isolated from Patients"

    Article Title: Characterization of H5N1 Influenza Virus Variants with Hemagglutinin Mutations Isolated from Patients

    Journal: mBio

    doi: 10.1128/mBio.00081-15

    Effects of HA mutations on viral replication in primary human airway cells. Small airway epithelial cells were infected with viruses at an MOI of 0.1. The culture supernatants were harvested at 24 (A and D), 48 (B and E), and 72 (C and F) h postinfection and assayed by quantitative real-time RT-PCR to determine the amount of progeny virus RNA. The amounts of progeny virus RNA produced by the HA mutant viruses (A, B, and C) and the seasonal human viruses (D, E, and F) are expressed relative to the amount produced by EG/D1 (wt) virus. Each data point is the mean ± SD from triplicate experiments. Colors on the x axis highlight the categories of HA mutations (A, B, and C) and virus strains (D, E, and F) as indicated. *, P
    Figure Legend Snippet: Effects of HA mutations on viral replication in primary human airway cells. Small airway epithelial cells were infected with viruses at an MOI of 0.1. The culture supernatants were harvested at 24 (A and D), 48 (B and E), and 72 (C and F) h postinfection and assayed by quantitative real-time RT-PCR to determine the amount of progeny virus RNA. The amounts of progeny virus RNA produced by the HA mutant viruses (A, B, and C) and the seasonal human viruses (D, E, and F) are expressed relative to the amount produced by EG/D1 (wt) virus. Each data point is the mean ± SD from triplicate experiments. Colors on the x axis highlight the categories of HA mutations (A, B, and C) and virus strains (D, E, and F) as indicated. *, P

    Techniques Used: Infection, Quantitative RT-PCR, Produced, Mutagenesis

    12) Product Images from "Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus"

    Article Title: Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus

    Journal: Veterinary Medicine and Science

    doi: 10.1002/vms3.18

    Viral excretion of experimentally inoculated ducks. The M viral RNA was detected by RRT ‐ PCR . Results are expressed as the M RNA copy number per ml. Vertical bars represent the results of individual inoculated Pekin ducks. Graphs represent tracheal
    Figure Legend Snippet: Viral excretion of experimentally inoculated ducks. The M viral RNA was detected by RRT ‐ PCR . Results are expressed as the M RNA copy number per ml. Vertical bars represent the results of individual inoculated Pekin ducks. Graphs represent tracheal

    Techniques Used: Quantitative RT-PCR

    13) Product Images from "Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus"

    Article Title: Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus

    Journal: Veterinary Medicine and Science

    doi: 10.1002/vms3.18

    Viral excretion of experimentally inoculated ducks. The M viral RNA was detected by RRT ‐ PCR . Results are expressed as the M RNA copy number per ml. Vertical bars represent the results of individual inoculated Pekin ducks. Graphs represent tracheal or cloacal excretion of virus by ducks inoculated with H7N1/chicken virus (a, b), H3N1/domestic duck virus (c, d) or H7N7/Canada goose virus (e, f). No swabs (tracheal or cloacal) were collected at 1 dpi for H3N1/domestic duck virus‐inoculated ducks.
    Figure Legend Snippet: Viral excretion of experimentally inoculated ducks. The M viral RNA was detected by RRT ‐ PCR . Results are expressed as the M RNA copy number per ml. Vertical bars represent the results of individual inoculated Pekin ducks. Graphs represent tracheal or cloacal excretion of virus by ducks inoculated with H7N1/chicken virus (a, b), H3N1/domestic duck virus (c, d) or H7N7/Canada goose virus (e, f). No swabs (tracheal or cloacal) were collected at 1 dpi for H3N1/domestic duck virus‐inoculated ducks.

    Techniques Used: Quantitative RT-PCR

    Related Articles

    Multiplex Assay:

    Article Title: A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses
    Article Snippet: .. In addition, the duplex real‐time PCR using Quantitect Multiplex Probe RT‐PCR kit (Qiagen) had similar sensitivity to single real‐time PCR procedures using Quantitect Probe RT‐PCR kit (Qiagen) in several probe–primer sets (Supplementary Fig. 1B). .. Gene Expression Image A gene expression image was produced with TreeView and Cluster software by Michael Eisen, University of California at Berkeley ( http://rana.lbl.gov/EisenSoftware.htm ) [Eisen et al., ].

    Amplification:

    Article Title: Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus
    Article Snippet: .. The Quantitect Probe RRT‐PCR kit (QiagenGmBH, Hilden, Germany) was used for amplification using a Biosystems 7500 Real‐Time PCR Cycler (Applied Biosystems, Lennik, Belgium). ..

    Infection:

    Article Title: Characterization of H5N1 Influenza Virus Variants with Hemagglutinin Mutations Isolated from Patients
    Article Snippet: .. Progeny virus RNA was extracted from the culture medium of infected cells with a QIAamp viral RNA minikit (Qiagen) and assayed by quantitative real-time reverse transcription (RT)-PCR (QuantiTect probe RT-PCR kit, Qiagen), using primers targeting the M gene as previously described ( ). ..

    Real-time Polymerase Chain Reaction:

    Article Title: Inactivation of Middle East respiratory syndrome‐coronavirus in human plasma using amotosalen and ultraviolet A light
    Article Snippet: .. Real‐time RT‐qPCR was performed in 96‐well plates on a fast real‐time PCR system (Model 7500, Applied Biosystems) using an RT‐PCR kit (QuantiTect Probe, Qiagen) according to the manufacturer's instructions in a total reaction volume of 25 μL. .. RNA transcript corresponding to the same target region was generated using a cDNA synthesis kit (Superscript RT III, Invitrogen) from a plasmid containing the MERS‐CoV N gene according to the manufacturer's instructions and used to generate a standard curve to estimate the viral RNA copy number in each sample as previously described., Each run included a positive viral template control and no‐template negative control.

    Article Title: Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus
    Article Snippet: .. The Quantitect Probe RRT‐PCR kit (QiagenGmBH, Hilden, Germany) was used for amplification using a Biosystems 7500 Real‐Time PCR Cycler (Applied Biosystems, Lennik, Belgium). ..

    Article Title: A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses
    Article Snippet: .. In addition, the duplex real‐time PCR using Quantitect Multiplex Probe RT‐PCR kit (Qiagen) had similar sensitivity to single real‐time PCR procedures using Quantitect Probe RT‐PCR kit (Qiagen) in several probe–primer sets (Supplementary Fig. 1B). .. Gene Expression Image A gene expression image was produced with TreeView and Cluster software by Michael Eisen, University of California at Berkeley ( http://rana.lbl.gov/EisenSoftware.htm ) [Eisen et al., ].

    Article Title: Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus
    Article Snippet: .. Extracted viral RNA was quantitated with probe-based qRT-PCR by using a qPCR system (MX3000P, Stratagene, La Jolla, CA) with a one-step qRT-PCR kit (QuantiTect Probe RT-PCR Kit, QIAGEN) in accordance with the manufacturer’s protocol. .. The P289-P290 amplicon of TV in the RNA-dependent RNA polymerase gene of caliciviruses ( , ) was determined in silico with DNASTAR software (Madison, WI) and experimentally confirmed (results not shown here).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis
    Article Snippet: .. Real-time quantitative RT-PCR Transcript levels of individual genes were assayed in frozen tissue specimens by quantitative real time (qRT)-PCR, using the QuantiTect™ Probe RT-PCR Kit (QIAGEN, Valencia, CA). .. For the PCR, 50 μl reactions were set up with 100 ng of RNA, 0.4 μM primer, 0.2 μM dual labeled probe, 0.5 μl of QuantiTect™ Reverse Transcriptase Mix and QuantiTect™ Probe RT-PCR Master Mix.

    Article Title: Inactivation of Middle East respiratory syndrome‐coronavirus in human plasma using amotosalen and ultraviolet A light
    Article Snippet: .. Real‐time RT‐qPCR was performed in 96‐well plates on a fast real‐time PCR system (Model 7500, Applied Biosystems) using an RT‐PCR kit (QuantiTect Probe, Qiagen) according to the manufacturer's instructions in a total reaction volume of 25 μL. .. RNA transcript corresponding to the same target region was generated using a cDNA synthesis kit (Superscript RT III, Invitrogen) from a plasmid containing the MERS‐CoV N gene according to the manufacturer's instructions and used to generate a standard curve to estimate the viral RNA copy number in each sample as previously described., Each run included a positive viral template control and no‐template negative control.

    Article Title: Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis
    Article Snippet: .. Real-time RT-PCR was used to determine the quantitative levels of SAP1–10 mRNA transcripts in RNA samples using the GeneAmp ABI 5700 and 7500 PCR machines (PE Applied Biosystems) and the QuantiTect TaqMan probe RT-PCR kit (Qiagen) according to the manufacturer’s instructions. .. Each reaction mixture consisted of 1× RT-PCR buffer, 4 mM MgCl2 , 600 nM forward and reverse primer, 200 nM TaqMan probe, HotStarTaq DNA polymerase, omniscript and sensiscript RT enzymes, and template RNA.

    Article Title: A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses
    Article Snippet: .. In addition, the duplex real‐time PCR using Quantitect Multiplex Probe RT‐PCR kit (Qiagen) had similar sensitivity to single real‐time PCR procedures using Quantitect Probe RT‐PCR kit (Qiagen) in several probe–primer sets (Supplementary Fig. 1B). .. Gene Expression Image A gene expression image was produced with TreeView and Cluster software by Michael Eisen, University of California at Berkeley ( http://rana.lbl.gov/EisenSoftware.htm ) [Eisen et al., ].

    Article Title: Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus
    Article Snippet: .. Extracted viral RNA was quantitated with probe-based qRT-PCR by using a qPCR system (MX3000P, Stratagene, La Jolla, CA) with a one-step qRT-PCR kit (QuantiTect Probe RT-PCR Kit, QIAGEN) in accordance with the manufacturer’s protocol. .. The P289-P290 amplicon of TV in the RNA-dependent RNA polymerase gene of caliciviruses ( , ) was determined in silico with DNASTAR software (Madison, WI) and experimentally confirmed (results not shown here).

    Article Title: Characterization of H5N1 Influenza Virus Variants with Hemagglutinin Mutations Isolated from Patients
    Article Snippet: .. Progeny virus RNA was extracted from the culture medium of infected cells with a QIAamp viral RNA minikit (Qiagen) and assayed by quantitative real-time reverse transcription (RT)-PCR (QuantiTect probe RT-PCR kit, Qiagen), using primers targeting the M gene as previously described ( ). ..

    Quantitative RT-PCR:

    Article Title: Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis
    Article Snippet: .. Real-time quantitative RT-PCR Transcript levels of individual genes were assayed in frozen tissue specimens by quantitative real time (qRT)-PCR, using the QuantiTect™ Probe RT-PCR Kit (QIAGEN, Valencia, CA). .. For the PCR, 50 μl reactions were set up with 100 ng of RNA, 0.4 μM primer, 0.2 μM dual labeled probe, 0.5 μl of QuantiTect™ Reverse Transcriptase Mix and QuantiTect™ Probe RT-PCR Master Mix.

    Article Title: Inactivation of Middle East respiratory syndrome‐coronavirus in human plasma using amotosalen and ultraviolet A light
    Article Snippet: .. Real‐time RT‐qPCR was performed in 96‐well plates on a fast real‐time PCR system (Model 7500, Applied Biosystems) using an RT‐PCR kit (QuantiTect Probe, Qiagen) according to the manufacturer's instructions in a total reaction volume of 25 μL. .. RNA transcript corresponding to the same target region was generated using a cDNA synthesis kit (Superscript RT III, Invitrogen) from a plasmid containing the MERS‐CoV N gene according to the manufacturer's instructions and used to generate a standard curve to estimate the viral RNA copy number in each sample as previously described., Each run included a positive viral template control and no‐template negative control.

    Article Title: Evaluation of the kinetics of anti‐ NP and anti‐ HA antibody after infection of Pekin ducks with low pathogenic avian influenza virus
    Article Snippet: .. The Quantitect Probe RRT‐PCR kit (QiagenGmBH, Hilden, Germany) was used for amplification using a Biosystems 7500 Real‐Time PCR Cycler (Applied Biosystems, Lennik, Belgium). ..

    Article Title: Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis
    Article Snippet: .. Real-time RT-PCR was used to determine the quantitative levels of SAP1–10 mRNA transcripts in RNA samples using the GeneAmp ABI 5700 and 7500 PCR machines (PE Applied Biosystems) and the QuantiTect TaqMan probe RT-PCR kit (Qiagen) according to the manufacturer’s instructions. .. Each reaction mixture consisted of 1× RT-PCR buffer, 4 mM MgCl2 , 600 nM forward and reverse primer, 200 nM TaqMan probe, HotStarTaq DNA polymerase, omniscript and sensiscript RT enzymes, and template RNA.

    Article Title: Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus
    Article Snippet: .. Extracted viral RNA was quantitated with probe-based qRT-PCR by using a qPCR system (MX3000P, Stratagene, La Jolla, CA) with a one-step qRT-PCR kit (QuantiTect Probe RT-PCR Kit, QIAGEN) in accordance with the manufacturer’s protocol. .. The P289-P290 amplicon of TV in the RNA-dependent RNA polymerase gene of caliciviruses ( , ) was determined in silico with DNASTAR software (Madison, WI) and experimentally confirmed (results not shown here).

    Polymerase Chain Reaction:

    Article Title: Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis
    Article Snippet: .. Real-time RT-PCR was used to determine the quantitative levels of SAP1–10 mRNA transcripts in RNA samples using the GeneAmp ABI 5700 and 7500 PCR machines (PE Applied Biosystems) and the QuantiTect TaqMan probe RT-PCR kit (Qiagen) according to the manufacturer’s instructions. .. Each reaction mixture consisted of 1× RT-PCR buffer, 4 mM MgCl2 , 600 nM forward and reverse primer, 200 nM TaqMan probe, HotStarTaq DNA polymerase, omniscript and sensiscript RT enzymes, and template RNA.

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    Qiagen quantitect taqman probe rt pcr kit
    Quantitative <t>TaqMan</t> <t>RT-PCR</t>
    Quantitect Taqman Probe Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quantitative TaqMan RT-PCR

    Journal:

    Article Title: Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis

    doi: 10.1099/mic.0.2008/022293-0

    Figure Lengend Snippet: Quantitative TaqMan RT-PCR

    Article Snippet: Real-time RT-PCR was used to determine the quantitative levels of SAP1–10 mRNA transcripts in RNA samples using the GeneAmp ABI 5700 and 7500 PCR machines (PE Applied Biosystems) and the QuantiTect TaqMan probe RT-PCR kit (Qiagen) according to the manufacturer’s instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction

    Establishment and validation of multivirus real‐time PCR. A : Procedure of the multivirus real‐time PCR system. DNA sample was mixed with Quantitect 2× master mix, and RNA sample was mixed with Quantitect 2× master mix and reverse transcriptase (RT) mix. These mixtures were poured into each well in a 96‐well plate at 10 µl per well. Ten microliters of 2× probe and primer mix were then added to each well in a premixed 96‐well plate. Finally, the virus genes were amplified and detected in a real‐time PCR machine for 2 hr. B : Validation of the multivirus real‐time PCR. A gene expression image by TreeView software based on the results of the multivirus real‐time PCR for control samples is shown. A horizontal line shows each probe–primer set and a vertical line is one sample of positive control. Gray vertical lines indicate no sample. A scale bar indicates copy number of color. A green box indicates specific reactions of target positive controls in specific probe–primer sets. Arrows of (a–c) also show specific signals. The arrow (a) shows positive signal for TTV in a brain sample with both JCV and TTV infection. The arrows (b1–3) show that a probe–primer set for pan‐enterovirus reacted with poliovirus (b1), Coxsackievirus B3 (b2), and Echovirus 6 (b3) positive samples. The arrow (c) shows that a probe–primer set for influenza virus A reacted with H5N1 influenza virus. Details of positive controls were listed in Supplementary Table II.

    Journal: Journal of Medical Virology

    Article Title: A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses

    doi: 10.1002/jmv.21962

    Figure Lengend Snippet: Establishment and validation of multivirus real‐time PCR. A : Procedure of the multivirus real‐time PCR system. DNA sample was mixed with Quantitect 2× master mix, and RNA sample was mixed with Quantitect 2× master mix and reverse transcriptase (RT) mix. These mixtures were poured into each well in a 96‐well plate at 10 µl per well. Ten microliters of 2× probe and primer mix were then added to each well in a premixed 96‐well plate. Finally, the virus genes were amplified and detected in a real‐time PCR machine for 2 hr. B : Validation of the multivirus real‐time PCR. A gene expression image by TreeView software based on the results of the multivirus real‐time PCR for control samples is shown. A horizontal line shows each probe–primer set and a vertical line is one sample of positive control. Gray vertical lines indicate no sample. A scale bar indicates copy number of color. A green box indicates specific reactions of target positive controls in specific probe–primer sets. Arrows of (a–c) also show specific signals. The arrow (a) shows positive signal for TTV in a brain sample with both JCV and TTV infection. The arrows (b1–3) show that a probe–primer set for pan‐enterovirus reacted with poliovirus (b1), Coxsackievirus B3 (b2), and Echovirus 6 (b3) positive samples. The arrow (c) shows that a probe–primer set for influenza virus A reacted with H5N1 influenza virus. Details of positive controls were listed in Supplementary Table II.

    Article Snippet: In addition, the duplex real‐time PCR using Quantitect Multiplex Probe RT‐PCR kit (Qiagen) had similar sensitivity to single real‐time PCR procedures using Quantitect Probe RT‐PCR kit (Qiagen) in several probe–primer sets (Supplementary Fig. 1B).

    Techniques: Real-time Polymerase Chain Reaction, Amplification, Expressing, Software, Positive Control, Infection

    qRT-PCR

    Journal: Journal of food protection

    Article Title: Inactivation of the Tulane Virus, a Novel Surrogate for the Human Norovirus

    doi: 10.4315/0362-028X.JFP-12-361

    Figure Lengend Snippet: qRT-PCR

    Article Snippet: Extracted viral RNA was quantitated with probe-based qRT-PCR by using a qPCR system (MX3000P, Stratagene, La Jolla, CA) with a one-step qRT-PCR kit (QuantiTect Probe RT-PCR Kit, QIAGEN) in accordance with the manufacturer’s protocol.

    Techniques: Quantitative RT-PCR

    Pharmacodynamic effects of TGF-β antagonists in vivo . A . Protein extracts from snap-frozen uninvolved lung tissue of mice treated with vehicle and LY2109761 and of mice treated with controls and 1D11 were prepared and subject to Western blotting using rabbit phosphoSmad2 antibody (1:1000 dilution). Whole cell lysate from SCP2TR was used as a control (Co). Treatment with both antagonists resulted in a reduction of phosphorylated Smad2 levels. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. B . RNA was extracted from snap-frozen uninvolved kidneys of mice treated with vehicle or LY2109761 using Trizol reagent (Invitrogen) and purified using RNeasy mini columns (Qiagen) according to the manufacturer's instructions. Transcript levels of CTGF and PAI-1 were assayed using the QuantiTect™ Probe RT-PCR Kit on a Mx4000 ® Multiplex Quantitative PCR System (Stratagene). Treatment with LY2109761 significantly reduced mRNA levels of both CTGF and PAI-1 relative to GAPDH mRNA. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. C . RNA was extracted from snap-frozen lungs of mice treated with vehicle, isotype control antibody or 1D11 and transcript levels determined as described above. No significant reduction in CTGF or PAI-1 mRNA levels could be detected in the 1D11-treated group. Values represent the means and SD of two independent experiments, 3 mice per group. Unpaired 2-sided t-test was used for comparisons.

    Journal: Molecular Cancer

    Article Title: Targeting the Transforming Growth Factor-? pathway inhibits human basal-like breast cancer metastasis

    doi: 10.1186/1476-4598-9-122

    Figure Lengend Snippet: Pharmacodynamic effects of TGF-β antagonists in vivo . A . Protein extracts from snap-frozen uninvolved lung tissue of mice treated with vehicle and LY2109761 and of mice treated with controls and 1D11 were prepared and subject to Western blotting using rabbit phosphoSmad2 antibody (1:1000 dilution). Whole cell lysate from SCP2TR was used as a control (Co). Treatment with both antagonists resulted in a reduction of phosphorylated Smad2 levels. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. B . RNA was extracted from snap-frozen uninvolved kidneys of mice treated with vehicle or LY2109761 using Trizol reagent (Invitrogen) and purified using RNeasy mini columns (Qiagen) according to the manufacturer's instructions. Transcript levels of CTGF and PAI-1 were assayed using the QuantiTect™ Probe RT-PCR Kit on a Mx4000 ® Multiplex Quantitative PCR System (Stratagene). Treatment with LY2109761 significantly reduced mRNA levels of both CTGF and PAI-1 relative to GAPDH mRNA. Values represent the means and SD of three mice per group. Unpaired 2-sided t-test was used for comparisons. C . RNA was extracted from snap-frozen lungs of mice treated with vehicle, isotype control antibody or 1D11 and transcript levels determined as described above. No significant reduction in CTGF or PAI-1 mRNA levels could be detected in the 1D11-treated group. Values represent the means and SD of two independent experiments, 3 mice per group. Unpaired 2-sided t-test was used for comparisons.

    Article Snippet: Real-time quantitative RT-PCR Transcript levels of individual genes were assayed in frozen tissue specimens by quantitative real time (qRT)-PCR, using the QuantiTect™ Probe RT-PCR Kit (QIAGEN, Valencia, CA).

    Techniques: In Vivo, Mouse Assay, Western Blot, Purification, Reverse Transcription Polymerase Chain Reaction, Multiplex Assay, Real-time Polymerase Chain Reaction