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    Thermo Fisher steponeplus realtime pcr system
    STING mutants fail to respond to DNA-adduct forming agents. The reconstituted primary Sting −/− MEF cells with empty vector (pBabe), hSTING, R169W, or P203S were treated DMBA (20 μg/ml) or cisplatin (10 μM) for 48 hr. Total RNA was extracted and <t>realtime</t> <t>PCR</t> was performed with the indicated probes after cDNA synthesis. Data shown here are the averages ± SD (n = 3). Asterisks indicate significant difference (p
    Steponeplus Realtime Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 13752 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    STING mutants fail to respond to DNA-adduct forming agents. The reconstituted primary Sting −/− MEF cells with empty vector (pBabe), hSTING, R169W, or P203S were treated DMBA (20 μg/ml) or cisplatin (10 μM) for 48 hr. Total RNA was extracted and realtime PCR was performed with the indicated probes after cDNA synthesis. Data shown here are the averages ± SD (n = 3). Asterisks indicate significant difference (p

    Journal: Oncogene

    Article Title: Suppression of STING Signaling through Epigenetic Silencing and Missense Mutation Impedes DNA-Damage Mediated Cytokine Production

    doi: 10.1038/s41388-017-0120-0

    Figure Lengend Snippet: STING mutants fail to respond to DNA-adduct forming agents. The reconstituted primary Sting −/− MEF cells with empty vector (pBabe), hSTING, R169W, or P203S were treated DMBA (20 μg/ml) or cisplatin (10 μM) for 48 hr. Total RNA was extracted and realtime PCR was performed with the indicated probes after cDNA synthesis. Data shown here are the averages ± SD (n = 3). Asterisks indicate significant difference (p

    Article Snippet: Realtime PCR was performed using StepOnePlus Real-Time PCR system (Applied Biosystems).

    Techniques: Plasmid Preparation, Polymerase Chain Reaction

    Effect of the removal of the histone N-terminal tails on the thermal stabilities of the nucleosomes. Each nucleosome, containing tlH2A, tlH2B, tlH3, or tlH4 (final concentration, 2.25 μM), was mixed with SYPRO Orange, and the sample temperature was increased by the StepOnePlus™ Real-Time PCR unit (Applied Biosystems). The temperature gradient was from 25 to 95 °C, in steps of 1 °C/min. Fluorescence signals of SYPRO Orange bound to the denatured histones were measured, and the thermal denaturing curves of the wt, tlH2A, tlH2B, tlH3, and tlH4 nucleosomes are shown in black, yellow, pink, blue, and green, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: FEBS Open Bio

    Article Title: Contribution of histone N-terminal tails to the structure and stability of nucleosomes

    doi: 10.1016/j.fob.2013.08.007

    Figure Lengend Snippet: Effect of the removal of the histone N-terminal tails on the thermal stabilities of the nucleosomes. Each nucleosome, containing tlH2A, tlH2B, tlH3, or tlH4 (final concentration, 2.25 μM), was mixed with SYPRO Orange, and the sample temperature was increased by the StepOnePlus™ Real-Time PCR unit (Applied Biosystems). The temperature gradient was from 25 to 95 °C, in steps of 1 °C/min. Fluorescence signals of SYPRO Orange bound to the denatured histones were measured, and the thermal denaturing curves of the wt, tlH2A, tlH2B, tlH3, and tlH4 nucleosomes are shown in black, yellow, pink, blue, and green, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: The sample temperature was increased by the StepOnePlus™ Real-Time PCR unit (Applied Biosystems), and the fluorescence signals were measured with this system.

    Techniques: Concentration Assay, Real-time Polymerase Chain Reaction, Fluorescence

    Glutamate-induced ERK activation stimulates the expression of both HSP70 and VRK3 to suppress its sustained activity that causes cell death. ( a ) The protein levels of HSP70 and VRK3 were positively correlated with ERK activity. ( b ) Blocking ERK phosphorylation using PD98059, a specific MEK/ERK inhibitor, delayed an increase in both HSP70 and VRK3 protein upon glutamate exposure. ( c , d ) Inhibition of ERK activation delayed mRNA expression of HSP70 ( c ) and VRK3 ( d ) following glutamate treatment. The mRNA levels of endogenous genes were detected by quantitative real-time PCR. Values are normalized to RPL32 and shown as mean ± s.d., n = 3. Student’s t-tests, **P

    Journal: Scientific Reports

    Article Title: VRK3-mediated nuclear localization of HSP70 prevents glutamate excitotoxicity-induced apoptosis and Aβ accumulation via enhancement of ERK phosphatase VHR activity

    doi: 10.1038/srep38452

    Figure Lengend Snippet: Glutamate-induced ERK activation stimulates the expression of both HSP70 and VRK3 to suppress its sustained activity that causes cell death. ( a ) The protein levels of HSP70 and VRK3 were positively correlated with ERK activity. ( b ) Blocking ERK phosphorylation using PD98059, a specific MEK/ERK inhibitor, delayed an increase in both HSP70 and VRK3 protein upon glutamate exposure. ( c , d ) Inhibition of ERK activation delayed mRNA expression of HSP70 ( c ) and VRK3 ( d ) following glutamate treatment. The mRNA levels of endogenous genes were detected by quantitative real-time PCR. Values are normalized to RPL32 and shown as mean ± s.d., n = 3. Student’s t-tests, **P

    Article Snippet: Quantitative real-time PCR The mRNA levels of endogenous genes were detected by quantitative real-time PCR using a StepOnePlus Real-Time PCR System (Applied Biosystems) with the FastStart Universal SYBR Green Master (Roche Applied Science).

    Techniques: Activation Assay, Expressing, Activity Assay, Blocking Assay, Inhibition, Real-time Polymerase Chain Reaction

    Representative qRT-PCR amplification and melting curve plots for the human IFN-γ- and CD8-specific amplicons. ( a ) CD8 and IFN-γ were amplified from cDNA of a normal HLA-A2+ human donor through qRT-PCR using the StepOnePlus™ Real-Time

    Journal: Methods in molecular biology (Clifton, N.J.)

    Article Title: Monitoring Antigen-Specific T Cell Responses Using Real-Time PCR

    doi: 10.1007/978-1-4939-1158-5_5

    Figure Lengend Snippet: Representative qRT-PCR amplification and melting curve plots for the human IFN-γ- and CD8-specific amplicons. ( a ) CD8 and IFN-γ were amplified from cDNA of a normal HLA-A2+ human donor through qRT-PCR using the StepOnePlus™ Real-Time

    Article Snippet: The following cycling parameters for PCR amplification are utilized on the StepOnePlus™ Real-Time PCR System (Life Technologies) (with melting curve analysis immediately following in order to confirm the absence of nonspecific amplification): 20 s at 95 °C.

    Techniques: Quantitative RT-PCR, Amplification

    qRT-PCR verification of selected differentially expressed genes . Gene expression in the seed and pericarp at 60 DAP was examined with a StepOnePlus™ Real-Time PCR Systems (Applied Biosystems) using SYBR Premix ExTaq™ (TaKaRa) according to the manufacturer's protocol. Relative expression values were calculated using the 2−ΔΔCt method by using EF1A gene as an internal control and were given as mean of the normalized expression levels of three replicates. For comparison, the gene expression level in the pericarp was arbitrarily set as 1.

    Journal: Frontiers in Plant Science

    Article Title: De novo Assembly and Characterization of the Fruit Transcriptome of Idesia polycarpa Reveals Candidate Genes for Lipid Biosynthesis

    doi: 10.3389/fpls.2016.00801

    Figure Lengend Snippet: qRT-PCR verification of selected differentially expressed genes . Gene expression in the seed and pericarp at 60 DAP was examined with a StepOnePlus™ Real-Time PCR Systems (Applied Biosystems) using SYBR Premix ExTaq™ (TaKaRa) according to the manufacturer's protocol. Relative expression values were calculated using the 2−ΔΔCt method by using EF1A gene as an internal control and were given as mean of the normalized expression levels of three replicates. For comparison, the gene expression level in the pericarp was arbitrarily set as 1.

    Article Snippet: The cDNA templates were then diluted 20-fold prior to use. qRT-PCR was performed with a StepOnePlus™ Real-Time PCR Systems (Applied Biosystems) using SYBR Premix ExTaq™ (TaKaRa) according to the manufacturer's protocol.

    Techniques: Quantitative RT-PCR, Expressing, Real-time Polymerase Chain Reaction