quantitative polymerase chain reaction qpcr kits  (Thermo Fisher)


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    Thermo Fisher quantitative polymerase chain reaction qpcr kits
    Quantitative Polymerase Chain Reaction Qpcr Kits, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative polymerase chain reaction qpcr kits/product/Thermo Fisher
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    quantitative polymerase chain reaction qpcr kits - by Bioz Stars, 2020-09
    92/100 stars

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    Real-time Polymerase Chain Reaction:

    Article Title: Paclitaxel Induce Apoptosis of Giant Cells Tumor of Bone via TP53INP1 Signaling
    Article Snippet: .. Puromycin, streptomycin and penicillin, Lipofectamine 3000 Transfection Reagent, RT reagent Kit, and quantitative polymerase chain reaction (qPCR) kits were from Thermo Fisher Scientific (Waltham, MA, USA). .. The CCK‐8 kit was provided by Dojindo Molecular Technologies (Kumamoto, Japan).

    Transfection:

    Article Title: Paclitaxel Induce Apoptosis of Giant Cells Tumor of Bone via TP53INP1 Signaling
    Article Snippet: .. Puromycin, streptomycin and penicillin, Lipofectamine 3000 Transfection Reagent, RT reagent Kit, and quantitative polymerase chain reaction (qPCR) kits were from Thermo Fisher Scientific (Waltham, MA, USA). .. The CCK‐8 kit was provided by Dojindo Molecular Technologies (Kumamoto, Japan).

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    Thermo Fisher sybr green i
    Correlation of detection of virus by Bio-Plex and RT-qPCR. Correlation between qPCR (Ct value) and Bio-Plex (MFI) detection of in-vitro grown ISAV standard curve by (A) <t>SYBR</t> <t>GREEN</t> I and (B)TaqMan; Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).
    Sybr Green I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 800 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green i/product/Thermo Fisher
    Average 90 stars, based on 800 article reviews
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    sybr green i - by Bioz Stars, 2020-09
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    84
    Thermo Fisher stabilized blood to ct nucleic acid preparation kit for qpcr
    An HSD activates peripheral nerve macrophages. a Representative flow cytometry dot plots showed an increase in the frequency of peripheral nerve macrophages (CD45 + CD11b + F4/80 + ) at 1 and 3 months of HSD feeding. b Histograms depicted the absolute macrophage (CD45 + CD11b + F4/80 + ) number, as well as those that were CD86 + macrophages. A quantitative analysis indicated a significant increase at 1 and 3 months of HSD feeding, and the number remained elevated after shifting HSD-fed mice to a ND, n = 6–9/group. c Histograms depicted the absolute macrophage (CD45 + CD11b + F4/80 + ) number as well as those that were CD206 + macrophages. An HSD favored the differentiation of CD86 + macrophages in the nerve more than those of CD206 + macrophages, n = 6–9/group. d A representative flow cytometry histogram showed the percentage of Ki67 + macrophages (CD45 + CD11b + F4/80 + ) in the sciatic nerves of HSD and ND fed mice. A quantitative analysis showed that HSD did not trigger significant macrophage proliferation in nerves, n = 6–8/group. e Longitudinal sections of mouse sciatic nerves were stained with CD16/32 (FcγII/III receptor), F4/80, and DAPI, showing an increased F4/80 + macrophage density in HSD nerves primarily colocalized with CD16/32. f Real-time quantitative <t>PCR</t> showed an elevated expression of IL-1β and IL-6 in the peripheral nerves at three months of HSD feeding, n = 6–10/group. Data were combined with from male and female mice. All data was presented as mean ± SEM and analyzed with an unpaired t test, ** p
    Stabilized Blood To Ct Nucleic Acid Preparation Kit For Qpcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stabilized blood to ct nucleic acid preparation kit for qpcr/product/Thermo Fisher
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    93
    Thermo Fisher ct one step rt qpcr kit
    <t>RNA</t> Immunoprecipitation experiments of confirmed BC200 targets. ( A ) <t>RT-qPCR</t> analysis of BC200, GAPDH and 7SL enrichment by immunoprecipitation of the indicated proteins. RNA extracted from 10% of the input sample was used as a reference to calculate percent of input for each RNA that was bound to the immunoprecipitated protein. Data represents the mean of three independent replicates ± standard deviation. Dashed line represents the threshold value of 5% input. ( B ) Percent input values of BC200 and 7SL were compared to GAPDH to demonstrate the degree of specificity of the interactions analyzed in (A). Dashed line represents the threshold value of 2-fold enrichment. ( C ) Immunoprecipitation efficiency was monitored by performing western blot on 50 μg of PRE and POST IP samples as well as 2% of the IP.
    Ct One Step Rt Qpcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Correlation of detection of virus by Bio-Plex and RT-qPCR. Correlation between qPCR (Ct value) and Bio-Plex (MFI) detection of in-vitro grown ISAV standard curve by (A) SYBR GREEN I and (B)TaqMan; Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).

    Journal: PLoS ONE

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology

    doi: 10.1371/journal.pone.0159155

    Figure Lengend Snippet: Correlation of detection of virus by Bio-Plex and RT-qPCR. Correlation between qPCR (Ct value) and Bio-Plex (MFI) detection of in-vitro grown ISAV standard curve by (A) SYBR GREEN I and (B)TaqMan; Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).

    Article Snippet: 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix.

    Techniques: Quantitative RT-PCR, Real-time Polymerase Chain Reaction, In Vitro, SYBR Green Assay

    RT-qPCR standard curves. Standard curve relating viral copy number to Ct value from RT-Polymerase chain reaction using 10-fold dilutions of in-vitro transcribed RNA using (a) SYBR ® Green I and (b) TaqMan chemistries.

    Journal: PLoS ONE

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology

    doi: 10.1371/journal.pone.0159155

    Figure Lengend Snippet: RT-qPCR standard curves. Standard curve relating viral copy number to Ct value from RT-Polymerase chain reaction using 10-fold dilutions of in-vitro transcribed RNA using (a) SYBR ® Green I and (b) TaqMan chemistries.

    Article Snippet: 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix.

    Techniques: Quantitative RT-PCR, Polymerase Chain Reaction, In Vitro, SYBR Green Assay

    An HSD activates peripheral nerve macrophages. a Representative flow cytometry dot plots showed an increase in the frequency of peripheral nerve macrophages (CD45 + CD11b + F4/80 + ) at 1 and 3 months of HSD feeding. b Histograms depicted the absolute macrophage (CD45 + CD11b + F4/80 + ) number, as well as those that were CD86 + macrophages. A quantitative analysis indicated a significant increase at 1 and 3 months of HSD feeding, and the number remained elevated after shifting HSD-fed mice to a ND, n = 6–9/group. c Histograms depicted the absolute macrophage (CD45 + CD11b + F4/80 + ) number as well as those that were CD206 + macrophages. An HSD favored the differentiation of CD86 + macrophages in the nerve more than those of CD206 + macrophages, n = 6–9/group. d A representative flow cytometry histogram showed the percentage of Ki67 + macrophages (CD45 + CD11b + F4/80 + ) in the sciatic nerves of HSD and ND fed mice. A quantitative analysis showed that HSD did not trigger significant macrophage proliferation in nerves, n = 6–8/group. e Longitudinal sections of mouse sciatic nerves were stained with CD16/32 (FcγII/III receptor), F4/80, and DAPI, showing an increased F4/80 + macrophage density in HSD nerves primarily colocalized with CD16/32. f Real-time quantitative PCR showed an elevated expression of IL-1β and IL-6 in the peripheral nerves at three months of HSD feeding, n = 6–10/group. Data were combined with from male and female mice. All data was presented as mean ± SEM and analyzed with an unpaired t test, ** p

    Journal: Journal of Neuroinflammation

    Article Title: High-salt diet decreases mechanical thresholds in mice that is mediated by a CCR2-dependent mechanism

    doi: 10.1186/s12974-020-01858-6

    Figure Lengend Snippet: An HSD activates peripheral nerve macrophages. a Representative flow cytometry dot plots showed an increase in the frequency of peripheral nerve macrophages (CD45 + CD11b + F4/80 + ) at 1 and 3 months of HSD feeding. b Histograms depicted the absolute macrophage (CD45 + CD11b + F4/80 + ) number, as well as those that were CD86 + macrophages. A quantitative analysis indicated a significant increase at 1 and 3 months of HSD feeding, and the number remained elevated after shifting HSD-fed mice to a ND, n = 6–9/group. c Histograms depicted the absolute macrophage (CD45 + CD11b + F4/80 + ) number as well as those that were CD206 + macrophages. An HSD favored the differentiation of CD86 + macrophages in the nerve more than those of CD206 + macrophages, n = 6–9/group. d A representative flow cytometry histogram showed the percentage of Ki67 + macrophages (CD45 + CD11b + F4/80 + ) in the sciatic nerves of HSD and ND fed mice. A quantitative analysis showed that HSD did not trigger significant macrophage proliferation in nerves, n = 6–8/group. e Longitudinal sections of mouse sciatic nerves were stained with CD16/32 (FcγII/III receptor), F4/80, and DAPI, showing an increased F4/80 + macrophage density in HSD nerves primarily colocalized with CD16/32. f Real-time quantitative PCR showed an elevated expression of IL-1β and IL-6 in the peripheral nerves at three months of HSD feeding, n = 6–10/group. Data were combined with from male and female mice. All data was presented as mean ± SEM and analyzed with an unpaired t test, ** p

    Article Snippet: RNA extraction and real-time quantitative PCR Whole blood was collected from the sub-mandibular vein plexus and lyzed by Ack Lysing Buffer.

    Techniques: Flow Cytometry, Mouse Assay, Staining, Real-time Polymerase Chain Reaction, Expressing

    Resistin stimulates VCAM-1 expression and monocyte adhesion to osteoarthritis synovial fibroblasts (OASFs). ( A – D ) OASFs were incubated with resistin (0, 1, 3, and 10 ng/mL) only or resistin at 10 ng/mL + VCAM-1 antibody for 24 h. THP-1 cells were subsequently added to OASFs for 1 h. The adherence of THP-1 cells to cultured OASFs was photographed under fluorescence microscopy (n = 4) ( A ) and quantified ( B ). VCAM-1 expression according to varying concentrations of resistin (0, 1, 3, or 10 ng/mL) was quantified by Western blotting (n = 3) ( C ) and RT-qPCR analysis (n = 4) ( D ). ( E ) IHC staining of VCAM-1 for normal (n = 10) and OA synovium (n = 10). ** p

    Journal: Cells

    Article Title: Resistin Enhances VCAM-1 Expression and Monocyte Adhesion in Human Osteoarthritis Synovial Fibroblasts by Inhibiting MiR-381 Expression through the PKC, p38, and JNK Signaling Pathways

    doi: 10.3390/cells9061369

    Figure Lengend Snippet: Resistin stimulates VCAM-1 expression and monocyte adhesion to osteoarthritis synovial fibroblasts (OASFs). ( A – D ) OASFs were incubated with resistin (0, 1, 3, and 10 ng/mL) only or resistin at 10 ng/mL + VCAM-1 antibody for 24 h. THP-1 cells were subsequently added to OASFs for 1 h. The adherence of THP-1 cells to cultured OASFs was photographed under fluorescence microscopy (n = 4) ( A ) and quantified ( B ). VCAM-1 expression according to varying concentrations of resistin (0, 1, 3, or 10 ng/mL) was quantified by Western blotting (n = 3) ( C ) and RT-qPCR analysis (n = 4) ( D ). ( E ) IHC staining of VCAM-1 for normal (n = 10) and OA synovium (n = 10). ** p

    Article Snippet: Real-Time Quantitative PCR Analysis Total RNA was extracted from OASFs by TRIzol; reverse transcription used 1 μg of total RNA transcribed into cDNA by oligo (dT) primers.

    Techniques: Expressing, Incubation, Cell Culture, Fluorescence, Microscopy, Western Blot, Quantitative RT-PCR, Immunohistochemistry, Staining

    Resistin promotes VCAM-1 expression and monocyte adhesion by suppressing miR-381 expression. ( A ) OASFs were incubated with resistin (0, 1, 3, or 10 ng/mL), and the extent of miR-381 expression was quantified by the RT-qPCR assay. ( B – D ) OASFs were transfected with miR-381 mimic or control siRNA, then stimulated with resistin. The extent of VCAM-1 transcription was examined by the RT-qPCR assay (n = 4) ( B ). THP-1 cells were subsequently added to OASFs for 1 h. The adherence of THP-1 cells to cultured OASFs was photographed by fluorescence microscopy and quantified (n = 4) ( C , D ). ( E ) Schematic demonstration of wt-VCAM-1 3′-UTR and mut-VCAM-1 3′-UTR. MiR-381 bound with wt-VCAM-1 3′-UTR but not mut-VCAM-1 3′-UTR. ( F ) OASFs were transfected with the wt-VCAM-1-3’-UTR plasmid with or without miR-381 mimic, then stimulated with resistin. Relative luciferase activity was quantified (n = 5). ( G , H ) OASFs were pretreated with inhibitors of PKC, JNK, and p38 (n = 4) ( G ) and their respective siRNAs (n = 5) ( H ), then incubated with resistin for 24 h. miR-381 expression levels were quantified by the RT-qPCR assay. ( I , J ) OASFs were treated with inhibitors of PKC, JNK, and p38 (n = 5) ( I ) or their respective siRNAs (n = 5) ( J ) and transfected with the wt-VCAM-1-3’-UTR plasmid before being incubated with resistin for 24 h. Relative luciferase activity was quantified. ( K ) OASFs were transfected with the wt-VCAM-1-3’-UTR plasmid or mut-VCAM-1-3’-UTR plasmid, then stimulated with resistin for 24 h. Relative luciferase activity was quantified (n = 5). ** p

    Journal: Cells

    Article Title: Resistin Enhances VCAM-1 Expression and Monocyte Adhesion in Human Osteoarthritis Synovial Fibroblasts by Inhibiting MiR-381 Expression through the PKC, p38, and JNK Signaling Pathways

    doi: 10.3390/cells9061369

    Figure Lengend Snippet: Resistin promotes VCAM-1 expression and monocyte adhesion by suppressing miR-381 expression. ( A ) OASFs were incubated with resistin (0, 1, 3, or 10 ng/mL), and the extent of miR-381 expression was quantified by the RT-qPCR assay. ( B – D ) OASFs were transfected with miR-381 mimic or control siRNA, then stimulated with resistin. The extent of VCAM-1 transcription was examined by the RT-qPCR assay (n = 4) ( B ). THP-1 cells were subsequently added to OASFs for 1 h. The adherence of THP-1 cells to cultured OASFs was photographed by fluorescence microscopy and quantified (n = 4) ( C , D ). ( E ) Schematic demonstration of wt-VCAM-1 3′-UTR and mut-VCAM-1 3′-UTR. MiR-381 bound with wt-VCAM-1 3′-UTR but not mut-VCAM-1 3′-UTR. ( F ) OASFs were transfected with the wt-VCAM-1-3’-UTR plasmid with or without miR-381 mimic, then stimulated with resistin. Relative luciferase activity was quantified (n = 5). ( G , H ) OASFs were pretreated with inhibitors of PKC, JNK, and p38 (n = 4) ( G ) and their respective siRNAs (n = 5) ( H ), then incubated with resistin for 24 h. miR-381 expression levels were quantified by the RT-qPCR assay. ( I , J ) OASFs were treated with inhibitors of PKC, JNK, and p38 (n = 5) ( I ) or their respective siRNAs (n = 5) ( J ) and transfected with the wt-VCAM-1-3’-UTR plasmid before being incubated with resistin for 24 h. Relative luciferase activity was quantified. ( K ) OASFs were transfected with the wt-VCAM-1-3’-UTR plasmid or mut-VCAM-1-3’-UTR plasmid, then stimulated with resistin for 24 h. Relative luciferase activity was quantified (n = 5). ** p

    Article Snippet: Real-Time Quantitative PCR Analysis Total RNA was extracted from OASFs by TRIzol; reverse transcription used 1 μg of total RNA transcribed into cDNA by oligo (dT) primers.

    Techniques: Expressing, Incubation, Quantitative RT-PCR, Transfection, Cell Culture, Fluorescence, Microscopy, Plasmid Preparation, Luciferase, Activity Assay

    The PKC pathway is involved in resistin-enhanced VCAM-1 expression and monocyte adhesion. ( A – E ) OASFs were pretreated with a PKC inhibitor (GF109203x), a specific PKCα/β inhibitor (Gö6976), or a PKCα siRNA, then incubated with varying concentrations of resistin for 24 h. The adherence of THP-1 cells to cultured OASFs was photographed by fluorescence microscopy ( A ) and quantified (n = 4) ( B , C ). The transcription levels of VCAM-1 were quantified by the RT-qPCR assay (n = 4) ( D , E ). The extent of phosphorylation of PKCα/β under resistin (10 ng/mL) stimulation (for 0, 15, 30, 60, or 120 min) was quantified by Western blotting (n = 3) ( F ). *** p

    Journal: Cells

    Article Title: Resistin Enhances VCAM-1 Expression and Monocyte Adhesion in Human Osteoarthritis Synovial Fibroblasts by Inhibiting MiR-381 Expression through the PKC, p38, and JNK Signaling Pathways

    doi: 10.3390/cells9061369

    Figure Lengend Snippet: The PKC pathway is involved in resistin-enhanced VCAM-1 expression and monocyte adhesion. ( A – E ) OASFs were pretreated with a PKC inhibitor (GF109203x), a specific PKCα/β inhibitor (Gö6976), or a PKCα siRNA, then incubated with varying concentrations of resistin for 24 h. The adherence of THP-1 cells to cultured OASFs was photographed by fluorescence microscopy ( A ) and quantified (n = 4) ( B , C ). The transcription levels of VCAM-1 were quantified by the RT-qPCR assay (n = 4) ( D , E ). The extent of phosphorylation of PKCα/β under resistin (10 ng/mL) stimulation (for 0, 15, 30, 60, or 120 min) was quantified by Western blotting (n = 3) ( F ). *** p

    Article Snippet: Real-Time Quantitative PCR Analysis Total RNA was extracted from OASFs by TRIzol; reverse transcription used 1 μg of total RNA transcribed into cDNA by oligo (dT) primers.

    Techniques: Expressing, Incubation, Cell Culture, Fluorescence, Microscopy, Quantitative RT-PCR, Western Blot

    The p38 and JNK pathways are involved in resistin-enhanced VCAM-1 synthesis and monocyte adhesion. ( A – E ) OASFs were pretreated with inhibitors of JNK (SP600125) and p38 (SB203580) and their respective siRNAs, then incubated with resistin for 24 h. The adherence of THP-1 cells to cultured OASFs was photographed by fluorescence microscopy ( A ) and quantified (n = 4) ( B , C ). The transcription levels of VCAM-1 were quantified by the RT-qPCR assay (n = 4) ( D , E ). The extent of phosphorylation of p38 and JNK under resistin (10 ng/mL) stimulation (for 0, 15, 30, 60, or 120 min) was quantified by Western blotting (n = 3) ( F ). ( G ) OASFs were pretreated with a PKC inhibitor (GF109203x) or a specific PKCα/β inhibitor (Gö6976), then incubated with resistin for 24 h. The extent of p38 and JNK phosphorylation was quantified by Western blotting (n = 3). *** p

    Journal: Cells

    Article Title: Resistin Enhances VCAM-1 Expression and Monocyte Adhesion in Human Osteoarthritis Synovial Fibroblasts by Inhibiting MiR-381 Expression through the PKC, p38, and JNK Signaling Pathways

    doi: 10.3390/cells9061369

    Figure Lengend Snippet: The p38 and JNK pathways are involved in resistin-enhanced VCAM-1 synthesis and monocyte adhesion. ( A – E ) OASFs were pretreated with inhibitors of JNK (SP600125) and p38 (SB203580) and their respective siRNAs, then incubated with resistin for 24 h. The adherence of THP-1 cells to cultured OASFs was photographed by fluorescence microscopy ( A ) and quantified (n = 4) ( B , C ). The transcription levels of VCAM-1 were quantified by the RT-qPCR assay (n = 4) ( D , E ). The extent of phosphorylation of p38 and JNK under resistin (10 ng/mL) stimulation (for 0, 15, 30, 60, or 120 min) was quantified by Western blotting (n = 3) ( F ). ( G ) OASFs were pretreated with a PKC inhibitor (GF109203x) or a specific PKCα/β inhibitor (Gö6976), then incubated with resistin for 24 h. The extent of p38 and JNK phosphorylation was quantified by Western blotting (n = 3). *** p

    Article Snippet: Real-Time Quantitative PCR Analysis Total RNA was extracted from OASFs by TRIzol; reverse transcription used 1 μg of total RNA transcribed into cDNA by oligo (dT) primers.

    Techniques: Incubation, Cell Culture, Fluorescence, Microscopy, Quantitative RT-PCR, Western Blot

    RNA Immunoprecipitation experiments of confirmed BC200 targets. ( A ) RT-qPCR analysis of BC200, GAPDH and 7SL enrichment by immunoprecipitation of the indicated proteins. RNA extracted from 10% of the input sample was used as a reference to calculate percent of input for each RNA that was bound to the immunoprecipitated protein. Data represents the mean of three independent replicates ± standard deviation. Dashed line represents the threshold value of 5% input. ( B ) Percent input values of BC200 and 7SL were compared to GAPDH to demonstrate the degree of specificity of the interactions analyzed in (A). Dashed line represents the threshold value of 2-fold enrichment. ( C ) Immunoprecipitation efficiency was monitored by performing western blot on 50 μg of PRE and POST IP samples as well as 2% of the IP.

    Journal: Nucleic Acids Research

    Article Title: Comprehensive analysis of the BC200 ribonucleoprotein reveals a reciprocal regulatory function with CSDE1/UNR

    doi: 10.1093/nar/gky860

    Figure Lengend Snippet: RNA Immunoprecipitation experiments of confirmed BC200 targets. ( A ) RT-qPCR analysis of BC200, GAPDH and 7SL enrichment by immunoprecipitation of the indicated proteins. RNA extracted from 10% of the input sample was used as a reference to calculate percent of input for each RNA that was bound to the immunoprecipitated protein. Data represents the mean of three independent replicates ± standard deviation. Dashed line represents the threshold value of 5% input. ( B ) Percent input values of BC200 and 7SL were compared to GAPDH to demonstrate the degree of specificity of the interactions analyzed in (A). Dashed line represents the threshold value of 2-fold enrichment. ( C ) Immunoprecipitation efficiency was monitored by performing western blot on 50 μg of PRE and POST IP samples as well as 2% of the IP.

    Article Snippet: For both native and crosslinking immunoprecipitations, RT-qPCR analysis was performed using an Applied Biosystems StepOnePlus instrument with the RNA to Ct One-step RT-qPCR kit (Thermo-Fisher Scientific).

    Techniques: Immunoprecipitation, Quantitative RT-PCR, Standard Deviation, Western Blot

    CSDE1 knock-down decreases stability of the BC200 RNA. ( A ) RT-qPCR analysis of BC200 expression following 72-hour CSDE1 or control knock-down and Actinomycin D treatment at T = 0. Indicated half-lives were calculated by fitting the data to a one-phase decay equation with GraphPad Prism software. Data represents the mean of three independent biological replicates measured in duplicate. ( B ) Relative total RNA quantities purified at the indicated time points from an equal number of cells to monitor total RNA decay. Data represents the mean of three independent replicates ± standard deviation. ( C ) MCF-7 cells were reverse transfected with control or CSDE1 siRNA and following 24 h were forward transfected with the plasmids containing BC200 under control of either the U6 snRNA promoter or endogenous BC200 promoter. Absolute expression levels of RNA from plasmid was calculated by RT-qPCR in parallel to a standard curve generated with purified BC200 RNA. Data represents the mean of three independent replicates ± standard deviation. ( D ) Relative CSDE1 mRNA expression was monitored by RT-qPCR analysis of the same RNA samples as used in (C).

    Journal: Nucleic Acids Research

    Article Title: Comprehensive analysis of the BC200 ribonucleoprotein reveals a reciprocal regulatory function with CSDE1/UNR

    doi: 10.1093/nar/gky860

    Figure Lengend Snippet: CSDE1 knock-down decreases stability of the BC200 RNA. ( A ) RT-qPCR analysis of BC200 expression following 72-hour CSDE1 or control knock-down and Actinomycin D treatment at T = 0. Indicated half-lives were calculated by fitting the data to a one-phase decay equation with GraphPad Prism software. Data represents the mean of three independent biological replicates measured in duplicate. ( B ) Relative total RNA quantities purified at the indicated time points from an equal number of cells to monitor total RNA decay. Data represents the mean of three independent replicates ± standard deviation. ( C ) MCF-7 cells were reverse transfected with control or CSDE1 siRNA and following 24 h were forward transfected with the plasmids containing BC200 under control of either the U6 snRNA promoter or endogenous BC200 promoter. Absolute expression levels of RNA from plasmid was calculated by RT-qPCR in parallel to a standard curve generated with purified BC200 RNA. Data represents the mean of three independent replicates ± standard deviation. ( D ) Relative CSDE1 mRNA expression was monitored by RT-qPCR analysis of the same RNA samples as used in (C).

    Article Snippet: For both native and crosslinking immunoprecipitations, RT-qPCR analysis was performed using an Applied Biosystems StepOnePlus instrument with the RNA to Ct One-step RT-qPCR kit (Thermo-Fisher Scientific).

    Techniques: Quantitative RT-PCR, Expressing, Software, Purification, Standard Deviation, Transfection, Plasmid Preparation, Generated

    BC200 and CSDE1 expression are mutually codependent. ( A ) RT-qPCR analysis of BC200 RNA expression following transfection with the indicated RNA interference oligonucleotides. Data represents the mean of three independent replicates ± standard deviation. ( B ) RT-qPCR analysis of CSDE1 mRNA expression following transfection with the indicated RNA interference oligonucleotides. Data represents the mean of three independent replicates ± standard deviation. ( C ) Western blot analysis of protein samples from a 72-hour knock-down time-course with the indicated RNA interference oligonucleotides and antibodies. Data is representative of three independent replicates. ( D ) Densitometry measurements of CSDE1 protein expression from (C) as well as replicate experiments. Data represents the mean of three independent replicates ± standard deviation

    Journal: Nucleic Acids Research

    Article Title: Comprehensive analysis of the BC200 ribonucleoprotein reveals a reciprocal regulatory function with CSDE1/UNR

    doi: 10.1093/nar/gky860

    Figure Lengend Snippet: BC200 and CSDE1 expression are mutually codependent. ( A ) RT-qPCR analysis of BC200 RNA expression following transfection with the indicated RNA interference oligonucleotides. Data represents the mean of three independent replicates ± standard deviation. ( B ) RT-qPCR analysis of CSDE1 mRNA expression following transfection with the indicated RNA interference oligonucleotides. Data represents the mean of three independent replicates ± standard deviation. ( C ) Western blot analysis of protein samples from a 72-hour knock-down time-course with the indicated RNA interference oligonucleotides and antibodies. Data is representative of three independent replicates. ( D ) Densitometry measurements of CSDE1 protein expression from (C) as well as replicate experiments. Data represents the mean of three independent replicates ± standard deviation

    Article Snippet: For both native and crosslinking immunoprecipitations, RT-qPCR analysis was performed using an Applied Biosystems StepOnePlus instrument with the RNA to Ct One-step RT-qPCR kit (Thermo-Fisher Scientific).

    Techniques: Expressing, Quantitative RT-PCR, RNA Expression, Transfection, Standard Deviation, Western Blot