7300 real time pcr system Search Results


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  • 99
    Thermo Fisher 7300 real time pcr system
    Real-time quantitative <t>RT-PCR</t> analysis of the snail hepatopancreas cathepsin B transcript in resistant (BS-90) and susceptible (NMRI and M-line) pre- and postexposure to miracidia. RT-PCR was performed with the cathepsin B gene-specific primers using RNA from individual snails after various times postexposure. Gene-specific primers for the housekeeping gene myoglobin were used to assess sample uniformity and to confirm that the template cDNA was used in equivalent amounts for each amplification reaction. RT-PCR reactions were performed using an Applied Biosystems <t>7300</t> Real-Time PCR System, and detection of the fluorescently labeled product was performed at the end of the amplification period. The -fold increase of gene expression was calculated by comparative Ct method as described in Materials and Methods. A value of > 1-fold increase was considered to be significant (1 = value for nonexposed snails).
    7300 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 31735 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi 7300 real time pcr system
    SG-PERT assay on <t>ABI</t> 7300 Real-Time <t>PCR</t> System. (A) Amplification curves of indicated amount of replication competent HIV-1 (NL4-3 strain) (ng p24/mL) obtained by SG-PERT on the ABI 7300 instrument. The horizontal line represents the threshold line used to calculate Cq values. (B) Correlation between input levels of replication-competent HIV-1 virus and Cq values obtained by SG-PERT on the ABI 7300 instrument. p24 antigen concentration in the undiluted samples (value “0” on x-axis) was 3,100 ng/mL.
    Abi 7300 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 8061 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi prism 7300 real time pcr system
    SG-PERT assay on <t>ABI</t> 7300 Real-Time <t>PCR</t> System. (A) Amplification curves of indicated amount of replication competent HIV-1 (NL4-3 strain) (ng p24/mL) obtained by SG-PERT on the ABI 7300 instrument. The horizontal line represents the threshold line used to calculate Cq values. (B) Correlation between input levels of replication-competent HIV-1 virus and Cq values obtained by SG-PERT on the ABI 7300 instrument. p24 antigen concentration in the undiluted samples (value “0” on x-axis) was 3,100 ng/mL.
    Abi Prism 7300 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 2288 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher real time pcr system
    Cpc attenuated α-MSH-stimulated melanogenesis and elevated the abundance of intracellular cAMP . Cells were pretreated with 20 nM α-MSH for 30 mins, and then treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Tyrosinase activity (black) and melanin content (grey) were measured. (B) The expression of tyrosinase was determined by immunoblotting analysis (black) and <t>RT-PCR</t> (grey), using β-actin and <t>GAPDH</t> as internal standards, respectively. (C) The cAMP concentration was measured by enzyme immunoassay at assigned time intervals (10, 30, 60 min) after Cpc treatment. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P
    Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22019 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher 7300 fast real time pcr system
    Cpc attenuated α-MSH-stimulated melanogenesis and elevated the abundance of intracellular cAMP . Cells were pretreated with 20 nM α-MSH for 30 mins, and then treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Tyrosinase activity (black) and melanin content (grey) were measured. (B) The expression of tyrosinase was determined by immunoblotting analysis (black) and <t>RT-PCR</t> (grey), using β-actin and <t>GAPDH</t> as internal standards, respectively. (C) The cAMP concentration was measured by enzyme immunoassay at assigned time intervals (10, 30, 60 min) after Cpc treatment. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P
    7300 Fast Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1055 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher quantitative real time pcr
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    Quantitative Real Time Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 45481 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher abi 7300 real time pcr machine
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    Abi 7300 Real Time Pcr Machine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher 7300 7500 real time pcr system
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    7300 7500 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 215 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher prism 7300 real time pcr system
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    Prism 7300 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher model 7300 real time pcr system
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    Model 7300 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 244 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher abi prism 7300 sequence detection system
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    Abi Prism 7300 Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 7780 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher abi 7300 system
    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, <t>IFN-γ,</t> IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative <t>PCR.</t> Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P
    Abi 7300 System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1780 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Real-time quantitative RT-PCR analysis of the snail hepatopancreas cathepsin B transcript in resistant (BS-90) and susceptible (NMRI and M-line) pre- and postexposure to miracidia. RT-PCR was performed with the cathepsin B gene-specific primers using RNA from individual snails after various times postexposure. Gene-specific primers for the housekeeping gene myoglobin were used to assess sample uniformity and to confirm that the template cDNA was used in equivalent amounts for each amplification reaction. RT-PCR reactions were performed using an Applied Biosystems 7300 Real-Time PCR System, and detection of the fluorescently labeled product was performed at the end of the amplification period. The -fold increase of gene expression was calculated by comparative Ct method as described in Materials and Methods. A value of > 1-fold increase was considered to be significant (1 = value for nonexposed snails).

    Journal: The Journal of parasitology

    Article Title: DIFFERENCES IN CYSTEINE PROTEASE ACTIVITY IN SCHISTOSOMA MANSONI-RESISTANT AND -SUSCEPTIBLE BIOMPHALARIA GLABRATA AND CHARACTERIZATION OF THE HEPATOPANCREAS CATHEPSIN B FULL-LENGTH cDNA

    doi: 10.1645/GE-1410R.1

    Figure Lengend Snippet: Real-time quantitative RT-PCR analysis of the snail hepatopancreas cathepsin B transcript in resistant (BS-90) and susceptible (NMRI and M-line) pre- and postexposure to miracidia. RT-PCR was performed with the cathepsin B gene-specific primers using RNA from individual snails after various times postexposure. Gene-specific primers for the housekeeping gene myoglobin were used to assess sample uniformity and to confirm that the template cDNA was used in equivalent amounts for each amplification reaction. RT-PCR reactions were performed using an Applied Biosystems 7300 Real-Time PCR System, and detection of the fluorescently labeled product was performed at the end of the amplification period. The -fold increase of gene expression was calculated by comparative Ct method as described in Materials and Methods. A value of > 1-fold increase was considered to be significant (1 = value for nonexposed snails).

    Article Snippet: The RT-PCR reactions were performed using an Applied Biosystems 7300 Real-Time PCR System (Applied Biosystems, Foster City, California).

    Techniques: Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Amplification, Real-time Polymerase Chain Reaction, Labeling, Expressing

    SG-PERT assay on ABI 7300 Real-Time PCR System. (A) Amplification curves of indicated amount of replication competent HIV-1 (NL4-3 strain) (ng p24/mL) obtained by SG-PERT on the ABI 7300 instrument. The horizontal line represents the threshold line used to calculate Cq values. (B) Correlation between input levels of replication-competent HIV-1 virus and Cq values obtained by SG-PERT on the ABI 7300 instrument. p24 antigen concentration in the undiluted samples (value “0” on x-axis) was 3,100 ng/mL.

    Journal: PLoS ONE

    Article Title: Quantification of Reverse Transcriptase Activity by Real-Time PCR as a Fast and Accurate Method for Titration of HIV, Lenti- and Retroviral Vectors

    doi: 10.1371/journal.pone.0050859

    Figure Lengend Snippet: SG-PERT assay on ABI 7300 Real-Time PCR System. (A) Amplification curves of indicated amount of replication competent HIV-1 (NL4-3 strain) (ng p24/mL) obtained by SG-PERT on the ABI 7300 instrument. The horizontal line represents the threshold line used to calculate Cq values. (B) Correlation between input levels of replication-competent HIV-1 virus and Cq values obtained by SG-PERT on the ABI 7300 instrument. p24 antigen concentration in the undiluted samples (value “0” on x-axis) was 3,100 ng/mL.

    Article Snippet: For SG-PERT on the ABI 7300 real-time PCR system (Applied Biosystems, Foster City, CA, USA), a reaction mix was made using the ROX-containing qPCR Core kit for SYBR Green I from Eurogentec (Catalog #RT-QP73-05).

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

    Cpc attenuated α-MSH-stimulated melanogenesis and elevated the abundance of intracellular cAMP . Cells were pretreated with 20 nM α-MSH for 30 mins, and then treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Tyrosinase activity (black) and melanin content (grey) were measured. (B) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. (C) The cAMP concentration was measured by enzyme immunoassay at assigned time intervals (10, 30, 60 min) after Cpc treatment. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Journal: Journal of Biomedical Science

    Article Title: Antimelanogenic effect of c-phycocyanin through modulation of tyrosinase expression by upregulation of ERK and downregulation of p38 MAPK signaling pathways

    doi: 10.1186/1423-0127-18-74

    Figure Lengend Snippet: Cpc attenuated α-MSH-stimulated melanogenesis and elevated the abundance of intracellular cAMP . Cells were pretreated with 20 nM α-MSH for 30 mins, and then treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Tyrosinase activity (black) and melanin content (grey) were measured. (B) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. (C) The cAMP concentration was measured by enzyme immunoassay at assigned time intervals (10, 30, 60 min) after Cpc treatment. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Article Snippet: Quantitative PCR Quantitative PCR (Q-PCR) was performed with reaction mixtures containing total RNA (100 ng), one-step RT-PCR Master Mix Reagents (Applied Biosystems, Foster City, CA, USA), and probes (MITF, GAPDH ) on 7300 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Effect of Cpc on cAMP/MAPK/ERK pathway and MITF expression at protein and mRNA levels . Immunoblot analysis was performed with cell extract proteins treated with (A) Cpc (0.1 mg/mL) at assigned time intervals for ERK1/2 (control (black); CPC-treated (grey)), and (B) different Cpc concentration (0.05, 0.1, 0.2 mg/mL) at 540 min for MEK. (C) Cell extract proteins at assigned time intervals treated with Cpc (0.1 mg/mL) were examined by Immunoblot analysis for MITF using β-actin as internal standards (control (black); CPC-treated (grey)). (D) Different levels of Cpc (0.05, 0.1, 0.2 mg/mL) treated MITF mRNA were analyzed by Q-PCR at 540 min. (E) Immunoblot analysis treated with Cpc (0.1 mg/mL), PD98059 (PD, 20 μM), and Cpc+PD at 72 hrs were performed for the evaluation of MITF and tyrosinase expression (MITF (black); tyrosinase (grey)). Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Journal: Journal of Biomedical Science

    Article Title: Antimelanogenic effect of c-phycocyanin through modulation of tyrosinase expression by upregulation of ERK and downregulation of p38 MAPK signaling pathways

    doi: 10.1186/1423-0127-18-74

    Figure Lengend Snippet: Effect of Cpc on cAMP/MAPK/ERK pathway and MITF expression at protein and mRNA levels . Immunoblot analysis was performed with cell extract proteins treated with (A) Cpc (0.1 mg/mL) at assigned time intervals for ERK1/2 (control (black); CPC-treated (grey)), and (B) different Cpc concentration (0.05, 0.1, 0.2 mg/mL) at 540 min for MEK. (C) Cell extract proteins at assigned time intervals treated with Cpc (0.1 mg/mL) were examined by Immunoblot analysis for MITF using β-actin as internal standards (control (black); CPC-treated (grey)). (D) Different levels of Cpc (0.05, 0.1, 0.2 mg/mL) treated MITF mRNA were analyzed by Q-PCR at 540 min. (E) Immunoblot analysis treated with Cpc (0.1 mg/mL), PD98059 (PD, 20 μM), and Cpc+PD at 72 hrs were performed for the evaluation of MITF and tyrosinase expression (MITF (black); tyrosinase (grey)). Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Article Snippet: Quantitative PCR Quantitative PCR (Q-PCR) was performed with reaction mixtures containing total RNA (100 ng), one-step RT-PCR Master Mix Reagents (Applied Biosystems, Foster City, CA, USA), and probes (MITF, GAPDH ) on 7300 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Expressing, Concentration Assay, Polymerase Chain Reaction

    Effect of Cpc on viability of B16F10 melanoma cell, tyrosinase activity and melanin contents . Cells were treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Cell viability was determined by MTT assay as described in Materials and Methods. (B) Tyrosinase activity (black) and melanin content (grey) were measured. (C) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Journal: Journal of Biomedical Science

    Article Title: Antimelanogenic effect of c-phycocyanin through modulation of tyrosinase expression by upregulation of ERK and downregulation of p38 MAPK signaling pathways

    doi: 10.1186/1423-0127-18-74

    Figure Lengend Snippet: Effect of Cpc on viability of B16F10 melanoma cell, tyrosinase activity and melanin contents . Cells were treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Cell viability was determined by MTT assay as described in Materials and Methods. (B) Tyrosinase activity (black) and melanin content (grey) were measured. (C) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Article Snippet: Quantitative PCR Quantitative PCR (Q-PCR) was performed with reaction mixtures containing total RNA (100 ng), one-step RT-PCR Master Mix Reagents (Applied Biosystems, Foster City, CA, USA), and probes (MITF, GAPDH ) on 7300 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Activity Assay, MTT Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

    Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, IFN-γ, IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P

    Journal: Infection and Immunity

    Article Title: Inhibition of Interleukin-22 Attenuates Bacterial Load and Organ Failure during Acute Polymicrobial Sepsis ▿

    doi: 10.1128/IAI.01564-06

    Figure Lengend Snippet: Expression of IL-22 and IL-22R during septic peritonitis. (A and B) Organs were removed from wild-type C57BL/6 mice before (0 h) and 3 h, 6 h, and 12 h after CASP. Organs were homogenized, and RNA was prepared. IL-22, IL-22R1, IL-10, IFN-γ, IL-10R2, IL-10R2, and IL-22BP levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four mice per group and time point. (C) Constitutive expression of IL-22BP mRNA was detected by reverse transcription-PCR in spleen 0 h and 3 h, respectively, after sepsis induction in three individual mice. (D) Spleens were removed from wild-type C57BL/6 mice before (0 h) and 6 h after CASP. T, B, and non-T/B cells were isolated by MACS and flow cytometry cell sorting, and RNA was prepared. IL-22 levels were determined by quantitative PCR. Expression levels are given relative to those before CASP as calibrator. Results are derived from four individual experiments. (E) Purity of B220- (left panel) or Thy1.2-positive (right panel) cell populations used for IL-22 quantitative PCR determinations after magnetic sorting. *, P

    Article Snippet: The expression of IL-22, IL-10, gamma interferon (IFN-γ), IL-22R1, IL-10R2, IL-10R2, and IL-22BP was analyzed using quantitative real-time PCR (ABI 7300 Real Time PCR System; Applied Biosystems, Foster City, CA) in liver, kidney, and spleen RNA samples from mice 0 h, 3 h, 6 h, and 12 h after sepsis induction.

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Derivative Assay, Polymerase Chain Reaction, Isolation, Magnetic Cell Separation, Flow Cytometry, Cytometry, FACS