glyceraldehyde 3 phosphate dehydrogenase gapdh  (Qiagen)


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    Structured Review

    Qiagen glyceraldehyde 3 phosphate dehydrogenase gapdh
    Gene expression of epithelial to mesenchymal transition (EMT) related markers. Relative ( A ) alpha smooth muscle actin ( α-SMA ), ( B ) fibronectin ( FN ) and ( C ) vimentin ( VIM ) expression evaluated by Real-time PCR in BE 63/3 cells treated or untreated with Everolimus (EVE) (5 and 100 nM) or TGF-β (20 ng/mL); expression values were normalized to <t>glyceraldehyde-3-phosphate</t> dehydrogenase <t>(GAPDH).</t> Mean ± S.D. (error bars) of three separate experiments performed in triplicate. * p
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "In Vitro Identification of New Transcriptomic and miRNomic Profiles Associated with Pulmonary Fibrosis Induced by High Doses Everolimus: Looking for New Pathogenetic Markers and Therapeutic Targets"

    Article Title: In Vitro Identification of New Transcriptomic and miRNomic Profiles Associated with Pulmonary Fibrosis Induced by High Doses Everolimus: Looking for New Pathogenetic Markers and Therapeutic Targets

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19041250

    Gene expression of epithelial to mesenchymal transition (EMT) related markers. Relative ( A ) alpha smooth muscle actin ( α-SMA ), ( B ) fibronectin ( FN ) and ( C ) vimentin ( VIM ) expression evaluated by Real-time PCR in BE 63/3 cells treated or untreated with Everolimus (EVE) (5 and 100 nM) or TGF-β (20 ng/mL); expression values were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Mean ± S.D. (error bars) of three separate experiments performed in triplicate. * p
    Figure Legend Snippet: Gene expression of epithelial to mesenchymal transition (EMT) related markers. Relative ( A ) alpha smooth muscle actin ( α-SMA ), ( B ) fibronectin ( FN ) and ( C ) vimentin ( VIM ) expression evaluated by Real-time PCR in BE 63/3 cells treated or untreated with Everolimus (EVE) (5 and 100 nM) or TGF-β (20 ng/mL); expression values were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Mean ± S.D. (error bars) of three separate experiments performed in triplicate. * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Related Articles

    Amplification:

    Article Title: In Vitro Identification of New Transcriptomic and miRNomic Profiles Associated with Pulmonary Fibrosis Induced by High Doses Everolimus: Looking for New Pathogenetic Markers and Therapeutic Targets
    Article Snippet: Real-time PCR amplification reactions were performed in duplicate via SYBR Green chemistry on CFX-connect (Bio-Rad, Hercules, CA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad). .. Primers for α-SMA, VIM, FN, MMP12, CTGF, CDH6, COL12A1, FAP, KAL1, LBH, PIM1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were obtained from Qiagen (QuantiTect Primer Assay, Hilden, Germany).

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion
    Article Snippet: .. We analyzed TNF-α (PPR06411F; Qiagen), IL-1β (PPR06480B; Qiagen), IL-6 (PPR06483B; Qiagen), inducible nitric oxide (iNOS) (PPR44835A; Qiagen), CX3CR1 (PPR06709A; Qiagen), CD206 (Mannose receptor: PPR65066A; Qiagen), TGF-β1 (pPPR06430B; Qiagen), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (PPR06557B; Qiagen). cDNA was amplified using Power SYBR Green PCR Master Mix with primers in an Applied Biosystems Step One Plus system (Thermo Fisher Scientific) at 95°C for 10 minutes, followed by 40 cycles of 15 seconds at 95°C and 1 minute at 60°C. .. A melting curve was generated to examine the specificity of the amplification.

    Positive Control:

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany). .. Relative expression of FcεRIα,-β and -γ was calculated according to the Pfaffl method as follows: ΔΔCT = ΔCt(sample)- ΔCt(reference), where ΔCt(sample) is the Ct value of the gene of interest normalized to the endogenous housekeeping genes GAPDH and beta-Actin and ΔCt(reference) is the sample with the lowest ΔCt value. cDNA from palatine tonsils was used as positive control and included in each experiment as inter-run calibrator.

    Synthesized:

    Article Title: Translating golden retriever muscular dystrophy microarray findings to novel biomarkers for cardiac/skeletal muscle function in Duchenne Muscular Dystrophy
    Article Snippet: .. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as an internal control. cDNA was synthesized from RNA using iScript (Bio-Rad Laboratories, Hercules, CA), and relative gene expression was assessed using Taq SYBR Green PCR (Bio-Rad Laboratories) in a Bio-Rad CFX instrument, as previously described ( ). .. Protein analyses Western blot analysis was performed as previously described ( ) using rabbit anti-BDNF (Aviva Systems Biology, San Diego, CA), anti-SPP1, Rockland Immunochemicals, Inc., Limerick, PA), or anti-p90 (Cell Signaling Technologies, Inc., Danvers, MA).

    Quantitative RT-PCR:

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions. ..

    Article Title: BET Bromodomain Proteins Brd2, Brd3 and Brd4 Selectively Regulate Metabolic Pathways in the Pancreatic β-Cell
    Article Snippet: .. RT-qPCR RNA from INS-1 cells was isolated using an RNeasy kit (Qiagen, Waltham, MA) and quantified as published [ ] using 7500 Fast Real-Time PCR System, Power SYBR Green PCR Master Mix (Life Technologies, Grand Island, NY) and Quantitech primers for rat insulin-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen). ..

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: Paragraph title: Real-time RT-PCR ... Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany).

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Paragraph title: miRNA microarray and RT-qPCR. ... The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: .. Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA). .. MCMV IE1 primer sequences (forward: 5'-TCA GCC ATC AAC TCT GCT ACC AAC-3' , reverse: 5'-ATC TGA AAC AGC CGT ATA TCA TCT TG-3' ) were purchased from Integrated DNA Technologies (IDT, Redwood City, CA).

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: The resulting cDNAs from whole splenic cells, enriched splenic CD4+ T cells, enriched splenic macrophages, enriched splenic Gr-1-expressing cells, and ocular tissues from all animal groups were subjected to quantitative real-time RT-PCR assay to determine the level of transcription specific for IL-17 (Sense: 5′ CCT GGC GGC TAC AGT GAA G 3′; Antisense: 5′ TTT GGA CAC GCT GAG CTT TG 3′) (Integrated DNA Technologies, Coralville, IA), IL-23 (Sense: 5′ GGT TGA GCG GAA T 3′; Antisense: 5′ AGG GAG TGG GAA C 3′) (Integrated DNA Technologies), IL-6 (Qiagen, Valencia, CA), and IL-10 (Qiagen). .. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control.

    Real-time Polymerase Chain Reaction:

    Article Title: Translating golden retriever muscular dystrophy microarray findings to novel biomarkers for cardiac/skeletal muscle function in Duchenne Muscular Dystrophy
    Article Snippet: Paragraph title: Quantitative PCR (qPCR) ... Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as an internal control. cDNA was synthesized from RNA using iScript (Bio-Rad Laboratories, Hercules, CA), and relative gene expression was assessed using Taq SYBR Green PCR (Bio-Rad Laboratories) in a Bio-Rad CFX instrument, as previously described ( ).

    Article Title: In Vitro Identification of New Transcriptomic and miRNomic Profiles Associated with Pulmonary Fibrosis Induced by High Doses Everolimus: Looking for New Pathogenetic Markers and Therapeutic Targets
    Article Snippet: Paragraph title: 4.5. Real-Time PCR ... Primers for α-SMA, VIM, FN, MMP12, CTGF, CDH6, COL12A1, FAP, KAL1, LBH, PIM1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were obtained from Qiagen (QuantiTect Primer Assay, Hilden, Germany).

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Detection and quantification of gene expression for each gene of interest were performed using specific primers and the SYBR green PCR master mix (Applied Biosystems, Foster City, CA), and products were detected and quantified using an ABI Prism 7500 real-time PCR instrument coupled with sequence detection system software (Applied Biosystems). .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Inflammatory breast cancer: Activation of the aryl hydrocarbon receptor and its target CYP1B1 correlates closely with Wnt5a/b-β-catenin signalling, the stem cell phenotype and disease progression
    Article Snippet: Paragraph title: Quantitative real-time PCR ... The data presented as relative gene expression of AHR and CYP1B1 were evaluated using the 2−ΔΔCt method after normalization to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH ) (Qiagen, CA, USA) (10 pmol\μL).

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion
    Article Snippet: Paragraph title: Quantitative Real-Time PCR ... We analyzed TNF-α (PPR06411F; Qiagen), IL-1β (PPR06480B; Qiagen), IL-6 (PPR06483B; Qiagen), inducible nitric oxide (iNOS) (PPR44835A; Qiagen), CX3CR1 (PPR06709A; Qiagen), CD206 (Mannose receptor: PPR65066A; Qiagen), TGF-β1 (pPPR06430B; Qiagen), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (PPR06557B; Qiagen). cDNA was amplified using Power SYBR Green PCR Master Mix with primers in an Applied Biosystems Step One Plus system (Thermo Fisher Scientific) at 95°C for 10 minutes, followed by 40 cycles of 15 seconds at 95°C and 1 minute at 60°C.

    Article Title: Hepatic alterations are accompanied by changes to bile acid transporter-expressing neurons in the hypothalamus after traumatic brain injury
    Article Snippet: .. Real-time PCR RNA was extracted from liver tissue (n = 3 sham & 3 FPI per time point) and real-time PCR was performed as previously described using commercially available primers against mouse ASBT, TGR5, OATP, NTCP, Shh, Ihh, Gli 1, Gli 2, Gli 3 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen, Frederick, MD). ..

    Article Title: BET Bromodomain Proteins Brd2, Brd3 and Brd4 Selectively Regulate Metabolic Pathways in the Pancreatic β-Cell
    Article Snippet: .. RT-qPCR RNA from INS-1 cells was isolated using an RNeasy kit (Qiagen, Waltham, MA) and quantified as published [ ] using 7500 Fast Real-Time PCR System, Power SYBR Green PCR Master Mix (Life Technologies, Grand Island, NY) and Quantitech primers for rat insulin-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen). ..

    Article Title: Role of CD73 in renal sympathetic neurotransmission in the mouse kidney
    Article Snippet: .. Using gene-specific primers for the adenosine A1 receptor (Qiagen, Gaithersburg, MD, catalog number QT00301119) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen, catalog number QT01658692) semi-quantitative real-time PCR was performed using an Applied Biosystems 7900HT Real-Time PCR System (Carlsbad, CA). .. There were four samples (all from separate mice) per genotype and each sample was run in duplicate for each primer pair tested.

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Real-time PCR was performed using a SYBR green PCR kit (Qiagen) or TaqMan 2× universal PCR master mix (Applied Biosystems, Foster City, CA) with gene-specific primers. .. The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: .. Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA). .. MCMV IE1 primer sequences (forward: 5'-TCA GCC ATC AAC TCT GCT ACC AAC-3' , reverse: 5'-ATC TGA AAC AGC CGT ATA TCA TCT TG-3' ) were purchased from Integrated DNA Technologies (IDT, Redwood City, CA).

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control. .. Transcription levels were determined utilizing 7500 Fast Real-Time PCR System (Applied Biosystems, Carlsbad, CA) and average threshold cycles (Ct) were determined using the 7500 Fast Real-Time PCR software.

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay. .. Fluoxetine Effects on Amygdala FAAH Activity To examine the effects of fluoxetine on amygdala FAAH activity, mice were treated with the drug for 21 days and killed the following day.

    Microarray:

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Paragraph title: miRNA microarray and RT-qPCR. ... The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Two Tailed Test:

    Article Title: Hepatic alterations are accompanied by changes to bile acid transporter-expressing neurons in the hypothalamus after traumatic brain injury
    Article Snippet: Real-time PCR RNA was extracted from liver tissue (n = 3 sham & 3 FPI per time point) and real-time PCR was performed as previously described using commercially available primers against mouse ASBT, TGR5, OATP, NTCP, Shh, Ihh, Gli 1, Gli 2, Gli 3 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen, Frederick, MD). .. It is pertinent to note that a two-tailed t test revealed a trend toward decreased GAPDH in the FPI group, compared to sham.

    Luciferase:

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: U343 cells were transfected with 100 nM luciferase (siCTL) or siQKI with Lipofectamine RNAi MAX (Invitrogen). .. The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Expressing:

    Article Title: Translating golden retriever muscular dystrophy microarray findings to novel biomarkers for cardiac/skeletal muscle function in Duchenne Muscular Dystrophy
    Article Snippet: .. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as an internal control. cDNA was synthesized from RNA using iScript (Bio-Rad Laboratories, Hercules, CA), and relative gene expression was assessed using Taq SYBR Green PCR (Bio-Rad Laboratories) in a Bio-Rad CFX instrument, as previously described ( ). .. Protein analyses Western blot analysis was performed as previously described ( ) using rabbit anti-BDNF (Aviva Systems Biology, San Diego, CA), anti-SPP1, Rockland Immunochemicals, Inc., Limerick, PA), or anti-p90 (Cell Signaling Technologies, Inc., Danvers, MA).

    Article Title: In Vitro Identification of New Transcriptomic and miRNomic Profiles Associated with Pulmonary Fibrosis Induced by High Doses Everolimus: Looking for New Pathogenetic Markers and Therapeutic Targets
    Article Snippet: Primers for α-SMA, VIM, FN, MMP12, CTGF, CDH6, COL12A1, FAP, KAL1, LBH, PIM1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were obtained from Qiagen (QuantiTect Primer Assay, Hilden, Germany). .. The comparative C t method (ΔΔ C t ) was used to quantify gene expression and the relative quantification was calculated as 2−ΔΔC t .

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Detection and quantification of gene expression for each gene of interest were performed using specific primers and the SYBR green PCR master mix (Applied Biosystems, Foster City, CA), and products were detected and quantified using an ABI Prism 7500 real-time PCR instrument coupled with sequence detection system software (Applied Biosystems). .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Inflammatory breast cancer: Activation of the aryl hydrocarbon receptor and its target CYP1B1 correlates closely with Wnt5a/b-β-catenin signalling, the stem cell phenotype and disease progression
    Article Snippet: .. The data presented as relative gene expression of AHR and CYP1B1 were evaluated using the 2−ΔΔCt method after normalization to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH ) (Qiagen, CA, USA) (10 pmol\μL). .. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting Tissue lysates were prepared from fresh breast tissues obtained during surgery, as described by El-Shinawi and colleagues .

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions. ..

    Article Title: BET Bromodomain Proteins Brd2, Brd3 and Brd4 Selectively Regulate Metabolic Pathways in the Pancreatic β-Cell
    Article Snippet: RT-qPCR RNA from INS-1 cells was isolated using an RNeasy kit (Qiagen, Waltham, MA) and quantified as published [ ] using 7500 Fast Real-Time PCR System, Power SYBR Green PCR Master Mix (Life Technologies, Grand Island, NY) and Quantitech primers for rat insulin-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen). .. Relative mRNA expression levels were determined using ΔCt values and were normalized to GAPDH.

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany). .. Relative expression of FcεRIα,-β and -γ was calculated according to the Pfaffl method as follows: ΔΔCT = ΔCt(sample)- ΔCt(reference), where ΔCt(sample) is the Ct value of the gene of interest normalized to the endogenous housekeeping genes GAPDH and beta-Actin and ΔCt(reference) is the sample with the lowest ΔCt value. cDNA from palatine tonsils was used as positive control and included in each experiment as inter-run calibrator.

    Article Title: Role of CD73 in renal sympathetic neurotransmission in the mouse kidney
    Article Snippet: Using gene-specific primers for the adenosine A1 receptor (Qiagen, Gaithersburg, MD, catalog number QT00301119) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen, catalog number QT01658692) semi-quantitative real-time PCR was performed using an Applied Biosystems 7900HT Real-Time PCR System (Carlsbad, CA). .. The duplicates were averaged and the A1 receptor mRNA expression was normalized for GAPDH levels.

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA). .. To determine gene expression changes in host cell-derived transcripts, ΔCT values of target gene mRNA in experimental wells were compared with control wells by the 2-ΔΔCt method, yielding a relative fold change in mRNA expression for each treatment group.

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay. .. Fluoxetine Effects on Amygdala FAAH Activity To examine the effects of fluoxetine on amygdala FAAH activity, mice were treated with the drug for 21 days and killed the following day.

    Transfection:

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Forty-eight hours after transfection, the cells were transfected a second time, and they were harvested at 72 h. Total RNA was isolated using miRNAeasy minikits (Qiagen). miRNA microarray analysis was performed and the data were analyzed by LC Sciences using 3 biological replicates for siCTL- and siQKI-transfected U343 cells. .. The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Paragraph title: Quantitative real-time reverse transcriptase PCR (RT-PCR) assay. ... Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: Paragraph title: RNA extraction and real-time reverse transcriptase polyacrylamide chain reaction (RT-PCR) ... Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA).

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: Paragraph title: 2.7. Quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay ... Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control.

    Generated:

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion
    Article Snippet: We analyzed TNF-α (PPR06411F; Qiagen), IL-1β (PPR06480B; Qiagen), IL-6 (PPR06483B; Qiagen), inducible nitric oxide (iNOS) (PPR44835A; Qiagen), CX3CR1 (PPR06709A; Qiagen), CD206 (Mannose receptor: PPR65066A; Qiagen), TGF-β1 (pPPR06430B; Qiagen), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (PPR06557B; Qiagen). cDNA was amplified using Power SYBR Green PCR Master Mix with primers in an Applied Biosystems Step One Plus system (Thermo Fisher Scientific) at 95°C for 10 minutes, followed by 40 cycles of 15 seconds at 95°C and 1 minute at 60°C. .. A melting curve was generated to examine the specificity of the amplification.

    Sequencing:

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Detection and quantification of gene expression for each gene of interest were performed using specific primers and the SYBR green PCR master mix (Applied Biosystems, Foster City, CA), and products were detected and quantified using an ABI Prism 7500 real-time PCR instrument coupled with sequence detection system software (Applied Biosystems). .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Injection:

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Whole MCMV-infected eyes and whole contralateral mock-infected eyes (controls) collected at 3, 6, and 10 days after subretinal injection were stored in RNAlater solution (Ambion, Austin, TX) at −70°C prior to analysis. .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Isolation:

    Article Title: BET Bromodomain Proteins Brd2, Brd3 and Brd4 Selectively Regulate Metabolic Pathways in the Pancreatic β-Cell
    Article Snippet: .. RT-qPCR RNA from INS-1 cells was isolated using an RNeasy kit (Qiagen, Waltham, MA) and quantified as published [ ] using 7500 Fast Real-Time PCR System, Power SYBR Green PCR Master Mix (Life Technologies, Grand Island, NY) and Quantitech primers for rat insulin-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen). ..

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: Real-time RT-PCR Total mRNA was isolated from mucosal specimens using the GenElute™ Total RNA MiniPrep Kit (Sigma-Aldrich, St. Louis, MO, USA) and subjected to reverse transcription with the iScript™ cDNA synthesis kit (Bio-Rad, Hercules, CA, USA) following the manufacturer’s instructions. .. Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany).

    Article Title: Role of CD73 in renal sympathetic neurotransmission in the mouse kidney
    Article Snippet: Real-time PCR for A1 receptor mRNA Total RNA was isolated from kidneys obtained from both CD73+/+ and −/− mice using Trizol (Life Technologies, Carlsbad, CA) according to the manufacturer's instructions. .. Using gene-specific primers for the adenosine A1 receptor (Qiagen, Gaithersburg, MD, catalog number QT00301119) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen, catalog number QT01658692) semi-quantitative real-time PCR was performed using an Applied Biosystems 7900HT Real-Time PCR System (Carlsbad, CA).

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Forty-eight hours after transfection, the cells were transfected a second time, and they were harvested at 72 h. Total RNA was isolated using miRNAeasy minikits (Qiagen). miRNA microarray analysis was performed and the data were analyzed by LC Sciences using 3 biological replicates for siCTL- and siQKI-transfected U343 cells. .. The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: Total RNA was isolated by chloroform extraction and purified over PureLink® RNA Mini Kit spin cartridge filters according to the manufacturer’s instructions (Ambion/ThermoFisher). .. Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA).

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: Total RNA was isolated with RNeasy Kit (Qiagen, Germantown, MD) followed by DNase I treatment (Invitrogen, Grand Island, NY, USA) to eliminate DNA, to purify RNA. .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay.

    Size-exclusion Chromatography:

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control. .. Parameters for each quantitative real-time RT-PCR assay cycle were 15 min at 95 °C, 15 sec at 94 °C, 31 sec at 55 °C, 35 sec at 72 °C for a total of 45 cycles.

    Purification:

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Total RNA was extracted from each eye homogenate using the PureLink total RNA purification system (Invitrogen Life Technologies) according to the manufacturer's instructions, and total RNA amounts for each sample were determined using a SmartSpec 3000 spectrometer (Bio-Rad Laboratories, Hercules, CA). .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Inflammatory breast cancer: Activation of the aryl hydrocarbon receptor and its target CYP1B1 correlates closely with Wnt5a/b-β-catenin signalling, the stem cell phenotype and disease progression
    Article Snippet: Quantitative real-time PCR Total RNA was extracted from fresh breast tissue and cells using a GeneJETTM RNA Purification Kit (Thermo Scientific, ON, Canada). .. The data presented as relative gene expression of AHR and CYP1B1 were evaluated using the 2−ΔΔCt method after normalization to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH ) (Qiagen, CA, USA) (10 pmol\μL).

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: Total RNA was isolated by chloroform extraction and purified over PureLink® RNA Mini Kit spin cartridge filters according to the manufacturer’s instructions (Ambion/ThermoFisher). .. Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA).

    Polymerase Chain Reaction:

    Article Title: Translating golden retriever muscular dystrophy microarray findings to novel biomarkers for cardiac/skeletal muscle function in Duchenne Muscular Dystrophy
    Article Snippet: .. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as an internal control. cDNA was synthesized from RNA using iScript (Bio-Rad Laboratories, Hercules, CA), and relative gene expression was assessed using Taq SYBR Green PCR (Bio-Rad Laboratories) in a Bio-Rad CFX instrument, as previously described ( ). .. Protein analyses Western blot analysis was performed as previously described ( ) using rabbit anti-BDNF (Aviva Systems Biology, San Diego, CA), anti-SPP1, Rockland Immunochemicals, Inc., Limerick, PA), or anti-p90 (Cell Signaling Technologies, Inc., Danvers, MA).

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Paragraph title: Quantitative real-time reverse transcriptase PCR (RT-PCR) assay. ... Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion
    Article Snippet: .. We analyzed TNF-α (PPR06411F; Qiagen), IL-1β (PPR06480B; Qiagen), IL-6 (PPR06483B; Qiagen), inducible nitric oxide (iNOS) (PPR44835A; Qiagen), CX3CR1 (PPR06709A; Qiagen), CD206 (Mannose receptor: PPR65066A; Qiagen), TGF-β1 (pPPR06430B; Qiagen), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (PPR06557B; Qiagen). cDNA was amplified using Power SYBR Green PCR Master Mix with primers in an Applied Biosystems Step One Plus system (Thermo Fisher Scientific) at 95°C for 10 minutes, followed by 40 cycles of 15 seconds at 95°C and 1 minute at 60°C. .. A melting curve was generated to examine the specificity of the amplification.

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions. ..

    Article Title: BET Bromodomain Proteins Brd2, Brd3 and Brd4 Selectively Regulate Metabolic Pathways in the Pancreatic β-Cell
    Article Snippet: .. RT-qPCR RNA from INS-1 cells was isolated using an RNeasy kit (Qiagen, Waltham, MA) and quantified as published [ ] using 7500 Fast Real-Time PCR System, Power SYBR Green PCR Master Mix (Life Technologies, Grand Island, NY) and Quantitech primers for rat insulin-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen). ..

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: The thermal profile of the PCR reaction was: initial denaturation at 95°C for 3 minutes, followed by 30 cycles of denaturation at 95°C for 10 seconds and annealing/elongation at 63°C for 30 seconds, and a final melting curve. .. Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany).

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Real-time PCR was performed using a SYBR green PCR kit (Qiagen) or TaqMan 2× universal PCR master mix (Applied Biosystems, Foster City, CA) with gene-specific primers. .. The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: Paragraph title: 2.7. Quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay ... Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control.

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay. .. Fluoxetine Effects on Amygdala FAAH Activity To examine the effects of fluoxetine on amygdala FAAH activity, mice were treated with the drug for 21 days and killed the following day.

    IA:

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: The resulting cDNAs from whole splenic cells, enriched splenic CD4+ T cells, enriched splenic macrophages, enriched splenic Gr-1-expressing cells, and ocular tissues from all animal groups were subjected to quantitative real-time RT-PCR assay to determine the level of transcription specific for IL-17 (Sense: 5′ CCT GGC GGC TAC AGT GAA G 3′; Antisense: 5′ TTT GGA CAC GCT GAG CTT TG 3′) (Integrated DNA Technologies, Coralville, IA), IL-23 (Sense: 5′ GGT TGA GCG GAA T 3′; Antisense: 5′ AGG GAG TGG GAA C 3′) (Integrated DNA Technologies), IL-6 (Qiagen, Valencia, CA), and IL-10 (Qiagen). .. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control.

    Activated Clotting Time Assay:

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: The following primers were used to amplify cDNA: FcεRI α: forward: 5′GAA ATG TGA CCT GCT GCT GA3′, reverse: 5′TGT GGC AGC TGG ACT ATG AG3′; beta-Actin: forward: 5′CTC TTC CAG CCT TCC TTC CT3′, reverse: 5′AGC ACT GTG TTG GCG TAC AG3′. .. Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany).

    Mouse Assay:

    Article Title: Role of CD73 in renal sympathetic neurotransmission in the mouse kidney
    Article Snippet: Real-time PCR for A1 receptor mRNA Total RNA was isolated from kidneys obtained from both CD73+/+ and −/− mice using Trizol (Life Technologies, Carlsbad, CA) according to the manufacturer's instructions. .. Using gene-specific primers for the adenosine A1 receptor (Qiagen, Gaithersburg, MD, catalog number QT00301119) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen, catalog number QT01658692) semi-quantitative real-time PCR was performed using an Applied Biosystems 7900HT Real-Time PCR System (Carlsbad, CA).

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: To explore possible effects of fluoxetine on amygdala eCB production and degradation, mice were treated with the drug for 21 days and killed the following day to remove the BLA. .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay.

    Chromatin Immunoprecipitation:

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: RNA quantity and integrity were evaluated with a 6000 Nano Chip™ Kit in an Agilent 2100 bioanalyzer (Agilent, Walbronn, Germany). .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions.

    Software:

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Detection and quantification of gene expression for each gene of interest were performed using specific primers and the SYBR green PCR master mix (Applied Biosystems, Foster City, CA), and products were detected and quantified using an ABI Prism 7500 real-time PCR instrument coupled with sequence detection system software (Applied Biosystems). .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions. ..

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: .. Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA). .. MCMV IE1 primer sequences (forward: 5'-TCA GCC ATC AAC TCT GCT ACC AAC-3' , reverse: 5'-ATC TGA AAC AGC CGT ATA TCA TCT TG-3' ) were purchased from Integrated DNA Technologies (IDT, Redwood City, CA).

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control. .. Transcription levels were determined utilizing 7500 Fast Real-Time PCR System (Applied Biosystems, Carlsbad, CA) and average threshold cycles (Ct) were determined using the 7500 Fast Real-Time PCR software.

    SYBR Green Assay:

    Article Title: Translating golden retriever muscular dystrophy microarray findings to novel biomarkers for cardiac/skeletal muscle function in Duchenne Muscular Dystrophy
    Article Snippet: .. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as an internal control. cDNA was synthesized from RNA using iScript (Bio-Rad Laboratories, Hercules, CA), and relative gene expression was assessed using Taq SYBR Green PCR (Bio-Rad Laboratories) in a Bio-Rad CFX instrument, as previously described ( ). .. Protein analyses Western blot analysis was performed as previously described ( ) using rabbit anti-BDNF (Aviva Systems Biology, San Diego, CA), anti-SPP1, Rockland Immunochemicals, Inc., Limerick, PA), or anti-p90 (Cell Signaling Technologies, Inc., Danvers, MA).

    Article Title: In Vitro Identification of New Transcriptomic and miRNomic Profiles Associated with Pulmonary Fibrosis Induced by High Doses Everolimus: Looking for New Pathogenetic Markers and Therapeutic Targets
    Article Snippet: Real-time PCR amplification reactions were performed in duplicate via SYBR Green chemistry on CFX-connect (Bio-Rad, Hercules, CA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad). .. Primers for α-SMA, VIM, FN, MMP12, CTGF, CDH6, COL12A1, FAP, KAL1, LBH, PIM1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were obtained from Qiagen (QuantiTect Primer Assay, Hilden, Germany).

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: Detection and quantification of gene expression for each gene of interest were performed using specific primers and the SYBR green PCR master mix (Applied Biosystems, Foster City, CA), and products were detected and quantified using an ABI Prism 7500 real-time PCR instrument coupled with sequence detection system software (Applied Biosystems). .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Inflammatory breast cancer: Activation of the aryl hydrocarbon receptor and its target CYP1B1 correlates closely with Wnt5a/b-β-catenin signalling, the stem cell phenotype and disease progression
    Article Snippet: Quantitative PCR was conducted as described previously by Ibrahim and colleagues via the StepOnePlus 96-well device (Applied Biosystems, CA, USA) in a 25 μL total reaction volume using 12.5 μL of SYBR green master mix (Applied Biosystems, Brumath, France), 1 μL of each primer (AHR , CYP1B1 , 10 pmol\μL (Qiagen, CA, USA); Wnt5a, 10 pmol\μL (a gift from Prof. Dr. Martin Götte, Department of Gynecology and Obstetrics, Münster University, Münster, Germany), Forward 5′-TCGTTAGCAGCATCAGTCCACA-3′ and reverse 5′-GACCTGTGCCTTCGTGCCTA-3′), 2.5 μL of cDNA and 8 μL of RNase free water. .. The data presented as relative gene expression of AHR and CYP1B1 were evaluated using the 2−ΔΔCt method after normalization to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH ) (Qiagen, CA, USA) (10 pmol\μL).

    Article Title: Mesenchymal Stem Cells Modulate the Functional Properties of Microglia via TGF-β Secretion
    Article Snippet: .. We analyzed TNF-α (PPR06411F; Qiagen), IL-1β (PPR06480B; Qiagen), IL-6 (PPR06483B; Qiagen), inducible nitric oxide (iNOS) (PPR44835A; Qiagen), CX3CR1 (PPR06709A; Qiagen), CD206 (Mannose receptor: PPR65066A; Qiagen), TGF-β1 (pPPR06430B; Qiagen), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (PPR06557B; Qiagen). cDNA was amplified using Power SYBR Green PCR Master Mix with primers in an Applied Biosystems Step One Plus system (Thermo Fisher Scientific) at 95°C for 10 minutes, followed by 40 cycles of 15 seconds at 95°C and 1 minute at 60°C. .. A melting curve was generated to examine the specificity of the amplification.

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions. ..

    Article Title: BET Bromodomain Proteins Brd2, Brd3 and Brd4 Selectively Regulate Metabolic Pathways in the Pancreatic β-Cell
    Article Snippet: .. RT-qPCR RNA from INS-1 cells was isolated using an RNeasy kit (Qiagen, Waltham, MA) and quantified as published [ ] using 7500 Fast Real-Time PCR System, Power SYBR Green PCR Master Mix (Life Technologies, Grand Island, NY) and Quantitech primers for rat insulin-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Qiagen). ..

    Article Title: Fc-Epsilon-RI, the High Affinity IgE-Receptor, Is Robustly Expressed in the Upper Gastrointestinal Tract and Modulated by Mucosal Inflammation
    Article Snippet: Quantification of mRNA was performed using the iQ SYBR Green Supermix (Bio-Rad) on a CFX96 Real time System (Bio-Rad). .. Primer sets for FcεRIβ and -γ subunits and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were purchased from Qiagen (Hilden, Germany).

    Article Title: The QKI-5 and QKI-6 RNA Binding Proteins Regulate the Expression of MicroRNA 7 in Glial Cells
    Article Snippet: Real-time PCR was performed using a SYBR green PCR kit (Qiagen) or TaqMan 2× universal PCR master mix (Applied Biosystems, Foster City, CA) with gene-specific primers. .. The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Qiagen (HS-GAPDH-1).

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: .. Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA). .. MCMV IE1 primer sequences (forward: 5'-TCA GCC ATC AAC TCT GCT ACC AAC-3' , reverse: 5'-ATC TGA AAC AGC CGT ATA TCA TCT TG-3' ) were purchased from Integrated DNA Technologies (IDT, Redwood City, CA).

    Article Title: Murine Cytomegalovirus Downregulates Interleukin-17 in Mice with Retrovirus-induced Immunosuppression that are Susceptible to Experimental Cytomegalovirus Retinitis
    Article Snippet: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen) served as the endogenous control. .. Briefly, 1.2 μl of cDNA was added to a reaction mixture of 15 μl of Power SYBR Green Master Mix (Applied Biosystems, Carlsbad, CA), 1.5 μl of forward and reverse primers for each gene, 9.9 μl of double-distilled water, and 0.9 μl of DMSO (Sigma, St. Louis, MO) for a total volume of 30 μl per reaction.

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay. .. Fluoxetine Effects on Amygdala FAAH Activity To examine the effects of fluoxetine on amygdala FAAH activity, mice were treated with the drug for 21 days and killed the following day.

    RNA Extraction:

    Article Title: Murine cytomegalovirus infection of mouse macrophages stimulates early expression of suppressor of cytokine signaling (SOCS)1 and SOCS3
    Article Snippet: Paragraph title: RNA extraction and real-time reverse transcriptase polyacrylamide chain reaction (RT-PCR) ... Real-time RT-PCR was performed using Applied Biosystems 7500 Fast Real-Time PCR System hardware and software with Power SYBR Green Master mix (Applied Biosystems, Foster City, CA) and with primer sets of mouse-specific SOCS1, SOCS3, SOCS5, IFN-α, IFN-β, IFN-γ, IL-6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) obtained from QIAgen (Valencia, CA).

    Article Title: Fluoxetine Facilitates Fear Extinction Through Amygdala Endocannabinoids
    Article Snippet: BLA punches were stored in RNAlater and kept in −20 °C until RNA extraction. .. Gene expression was quantified with QuantiTect Primer Assay and Power SYBR Green PCR master mix (Applied Biosystems, Grand Island, NY) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems) and normalized to the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (Gapdh ) (Qiagen; cat. no.: QT01658692, lot. no.: 176907735) using the QuantiTect Primer Assay.

    Selection:

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions. ..

    Concentration Assay:

    Article Title: Evidence For Multiple Cell Death Pathways during Development of Experimental Cytomegalovirus Retinitis in Mice with Retrovirus-Induced Immunosuppression: Apoptosis, Necroptosis, and Pyroptosis
    Article Snippet: After the total RNA concentration was normalized for each sample, cDNA synthesis was performed using the SuperScript III first-strand synthesis system (Invitrogen) according to the manufacturer's instructions. .. Primers used for detection and quantification of transcripts for TNF-α, TNFR1, TNFR2, active (cleaved) caspase 8, active (cleaved) caspase 3, TRAIL, TRAIL-R(DR5), Fas, FasL, caspase 9, cytochrome c , apoptotic protease-activating factor 1 (Apaf-1), receptor-interacting protein 1 (RIP1), RIP3, interleukin 1β (IL-1β), IL-6, IL-18, caspase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Qiagen Inc., Valencia, CA.

    Article Title: Inflammatory breast cancer: Activation of the aryl hydrocarbon receptor and its target CYP1B1 correlates closely with Wnt5a/b-β-catenin signalling, the stem cell phenotype and disease progression
    Article Snippet: The total RNA concentration was assessed by Infinite®200 PRO NanoQuant (Tecan, Zürich, Switzerland). .. The data presented as relative gene expression of AHR and CYP1B1 were evaluated using the 2−ΔΔCt method after normalization to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH ) (Qiagen, CA, USA) (10 pmol\μL).

    Lysis:

    Article Title: Subcutaneous immunotherapy induces alterations in monocytes and dendritic cells homeostasis in allergic rhinitis patients
    Article Snippet: Evaluation of IFNα mRNA expression by qRT-PCR Cell suspensions were centrifuged for 5 min at 300 g and the pellet was resuspended in 350 µl of RLT Lysis Buffer (Qiagen, Hilden, Germany). .. To select optimal housekeeping genes, normalization of gene expression was executed with geNorm Housekeeping Gene Selection Human Kit (Primer Design, Southampton, UK) and geNorm™ software (Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium). qRT-PCR was done with QuantiTect SYBR Green PCR Kit Gene expression, using optimized primers for IFNα and endogenous controls as beta-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Qiagen), according to the manufacturer’s instructions.

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  • 95
    Qiagen glyceraldehyde 3 phosphate dehydrogenase
    Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in the choroid plexus. Rabbit pups with confirmed IVH, injected intraventricularly with Hp (grey bars; n = 6) or sham (white bars; n = 6) 6 hours after confirmation of the bleeding, were euthanized at 24 hours of age and the brains were removed from the skulls and the choroid plexus was carefully removed from the lateral ventricles, snap frozen, and the mRNA expression of IL1R1 (A) , FAS (B) , NF-Κβ (C) , MCP-1 (D) , IL-8 (E) , IL-1β (F) , TNFα (G) , IL-6 (H) and HO-1 (I) were subsequently analyzed with real-time PCR, as described in Materials and Methods. mRNA expression for respective gene was normalized against those of <t>glyceraldehyde-3-phosphate</t> dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against samples from control pups. Results are presented as box plots displaying medians and 25 th and 75 th percentiles. Differences between IVH + Hp versus IVH + Sham at 24 hours were analyzed using the Mann-Whitney U -test. * P
    Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Qiagen human glyceraldehyde 3 phosphate dehydrogenase
    BAY 11-7082 reduced the acidic bile-induced gene expression profiling of NF-κB signaling pathway in treated normal human hypopharyngeal cells Heat maps were obtained by RT 2 -Profiler PCR array analysis for NF-κB signaling. The genes were clustered based on their biological role. The heat maps demonstrated the effect of BAY 11-7082 in gene expression of NF-κB pathway and NF-κB responsive genes (red color for maximum expression and green for minimum). Group 1: Acidic bile-treated cells with BAY 11-7082; Control Group: Acidic bile-treated cells without BAY 11-7082. (Gene expression has been normalized to two housekeeping genes; hGAPDH, human <t>glyceraldehyde-3-phosphate</t> dehydrogenase and RPLP0, ribosomal protein lateral stalk subunit P0).
    Human Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Qiagen, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Qiagen mouse glyceraldehyde 3 phosphate dehydrogenase gapdh cdna measurements
    mRNA expression levels of the target gene in various organs . mRNA levels were evaluated using real time RT-PCR. Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice. Mice were sacrificed at the indicated time points, and total RNA was extracted from the organs. After preparation of <t>cDNA,</t> PCR amplification of luciferase and <t>GAPDH</t> genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.
    Mouse Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Cdna Measurements, supplied by Qiagen, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse glyceraldehyde 3 phosphate dehydrogenase gapdh cdna measurements/product/Qiagen
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse glyceraldehyde 3 phosphate dehydrogenase gapdh cdna measurements - by Bioz Stars, 2020-02
    78/100 stars
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    Image Search Results


    Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in the choroid plexus. Rabbit pups with confirmed IVH, injected intraventricularly with Hp (grey bars; n = 6) or sham (white bars; n = 6) 6 hours after confirmation of the bleeding, were euthanized at 24 hours of age and the brains were removed from the skulls and the choroid plexus was carefully removed from the lateral ventricles, snap frozen, and the mRNA expression of IL1R1 (A) , FAS (B) , NF-Κβ (C) , MCP-1 (D) , IL-8 (E) , IL-1β (F) , TNFα (G) , IL-6 (H) and HO-1 (I) were subsequently analyzed with real-time PCR, as described in Materials and Methods. mRNA expression for respective gene was normalized against those of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against samples from control pups. Results are presented as box plots displaying medians and 25 th and 75 th percentiles. Differences between IVH + Hp versus IVH + Sham at 24 hours were analyzed using the Mann-Whitney U -test. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in the choroid plexus. Rabbit pups with confirmed IVH, injected intraventricularly with Hp (grey bars; n = 6) or sham (white bars; n = 6) 6 hours after confirmation of the bleeding, were euthanized at 24 hours of age and the brains were removed from the skulls and the choroid plexus was carefully removed from the lateral ventricles, snap frozen, and the mRNA expression of IL1R1 (A) , FAS (B) , NF-Κβ (C) , MCP-1 (D) , IL-8 (E) , IL-1β (F) , TNFα (G) , IL-6 (H) and HO-1 (I) were subsequently analyzed with real-time PCR, as described in Materials and Methods. mRNA expression for respective gene was normalized against those of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against samples from control pups. Results are presented as box plots displaying medians and 25 th and 75 th percentiles. Differences between IVH + Hp versus IVH + Sham at 24 hours were analyzed using the Mann-Whitney U -test. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Injection, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY

    Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10% (of cell culture medium volume) cerebrospinal fluid (CSF) from preterm infants with IVH or 10 μM metHb with or without the addition of 1.0 mg/ml Hp, for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10% (of cell culture medium volume) cerebrospinal fluid (CSF) from preterm infants with IVH or 10 μM metHb with or without the addition of 1.0 mg/ml Hp, for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Expressing, Cell Culture, Real-time Polymerase Chain Reaction

    CSF-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to culture medium containing 1 to 30% (of cell culture medium volume) CSF from preterm infants with intraventricular hemorrhage (IVH) for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: CSF-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to culture medium containing 1 to 30% (of cell culture medium volume) CSF from preterm infants with intraventricular hemorrhage (IVH) for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Expressing, Cell Culture, Real-time Polymerase Chain Reaction

    MetHb- and heme-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10 μM metHb or heme for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: MetHb- and heme-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10 μM metHb or heme for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

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

    Real-time PCR gene expression comparison between BM testes and germ cell-derived colonies. Relative gene expressions were compared between GDCs and 4-month-old BM testes.  GAPDH , glyceraldehyde 3-phosphate dehydrogenase;  PGP9.5 , protein gene product 9.5 (alternate designation: ubiquitin carboxy-terminal hydrolase L1);  GFRα-1 , glial cell line derived neurotrophic factor family receptor alpha 1;  PLZF , promyelocytic leukaemia zinc finger;  Oct4 , octamer-binding protein 4;  Nanog , homeobox transcription factor Nanog;  GATA4 , GATA-binding protein 4. (* P

    Journal: Scientific Reports

    Article Title: Vitrified canine testicular cells allow the formation of spermatogonial stem cells and seminiferous tubules following their xenotransplantation into nude mice

    doi: 10.1038/srep21919

    Figure Lengend Snippet: Real-time PCR gene expression comparison between BM testes and germ cell-derived colonies. Relative gene expressions were compared between GDCs and 4-month-old BM testes. GAPDH , glyceraldehyde 3-phosphate dehydrogenase; PGP9.5 , protein gene product 9.5 (alternate designation: ubiquitin carboxy-terminal hydrolase L1); GFRα-1 , glial cell line derived neurotrophic factor family receptor alpha 1; PLZF , promyelocytic leukaemia zinc finger; Oct4 , octamer-binding protein 4; Nanog , homeobox transcription factor Nanog; GATA4 , GATA-binding protein 4. (* P

    Article Snippet: Real-time PCR was performed with glyceraldehyde 3-phosphate dehydrogenase (GAPDH ), GFRα-1 , PLZF , Oct4 , Nanog , and GATA-binding protein 4 (GATA4 ) primers, using Rotor-Gene Q (Qiagen, Venlo, the Netherlands).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Derivative Assay, Binding Assay

    BAY 11-7082 reduced the acidic bile-induced gene expression profiling of NF-κB signaling pathway in treated normal human hypopharyngeal cells Heat maps were obtained by RT 2 -Profiler PCR array analysis for NF-κB signaling. The genes were clustered based on their biological role. The heat maps demonstrated the effect of BAY 11-7082 in gene expression of NF-κB pathway and NF-κB responsive genes (red color for maximum expression and green for minimum). Group 1: Acidic bile-treated cells with BAY 11-7082; Control Group: Acidic bile-treated cells without BAY 11-7082. (Gene expression has been normalized to two housekeeping genes; hGAPDH, human glyceraldehyde-3-phosphate dehydrogenase and RPLP0, ribosomal protein lateral stalk subunit P0).

    Journal: Oncotarget

    Article Title: Inhibition of NF-κB prevents the acidic bile-induced oncogenic mRNA phenotype, in human hypopharyngeal cells

    doi: 10.18632/oncotarget.23143

    Figure Lengend Snippet: BAY 11-7082 reduced the acidic bile-induced gene expression profiling of NF-κB signaling pathway in treated normal human hypopharyngeal cells Heat maps were obtained by RT 2 -Profiler PCR array analysis for NF-κB signaling. The genes were clustered based on their biological role. The heat maps demonstrated the effect of BAY 11-7082 in gene expression of NF-κB pathway and NF-κB responsive genes (red color for maximum expression and green for minimum). Group 1: Acidic bile-treated cells with BAY 11-7082; Control Group: Acidic bile-treated cells without BAY 11-7082. (Gene expression has been normalized to two housekeeping genes; hGAPDH, human glyceraldehyde-3-phosphate dehydrogenase and RPLP0, ribosomal protein lateral stalk subunit P0).

    Article Snippet: We performed reverse transcription (iScript cDNA synthesis kit; Bio-Rad) and real time qPCR analysis (Bio-Rad real time thermal cycler CFX96TM; Bio-Rad) using specific primers for target genes and reference housekeeping gene, human glyceraldehyde 3-phosphate dehydrogenase (h GAPDH) ( ; see online), (QuantiTect Primers Assays; Qiagen), and iQ™ SYBR Green Supermix (Bio-Rad).

    Techniques: Expressing, Polymerase Chain Reaction

    mRNA expression levels of the target gene in various organs . mRNA levels were evaluated using real time RT-PCR. Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice. Mice were sacrificed at the indicated time points, and total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Journal: Genetic Vaccines and Therapy

    Article Title: Biodistribution and blood clearance of plasmid DNA administered in arginine peptide complexes

    doi: 10.1186/1479-0556-9-13

    Figure Lengend Snippet: mRNA expression levels of the target gene in various organs . mRNA levels were evaluated using real time RT-PCR. Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice. Mice were sacrificed at the indicated time points, and total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Article Snippet: The thermal cycler protocol was set as follows: pre-incubation at 95°C for 10 s, amplification at 40 cycles at 95°C for 5 s, and 60°C for 40 s. For the mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA measurements, each sample was prepared following the manufacturer's instructions with a GAPDH primer set (Qiagen).

    Techniques: Expressing, Quantitative RT-PCR, Plasmid Preparation, Mouse Assay, Polymerase Chain Reaction, Amplification, Luciferase

    Effects of DNA dose on plasmid DNA expression after delivery in arginine/DNA complexes . Various amounts of plasmid DNA complexed with arginine peptide at an N/P ratio of 3:1 were intraperitoneally administered to mice, and mRNA levels were evaluated using real time RT-PCR. Total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Journal: Genetic Vaccines and Therapy

    Article Title: Biodistribution and blood clearance of plasmid DNA administered in arginine peptide complexes

    doi: 10.1186/1479-0556-9-13

    Figure Lengend Snippet: Effects of DNA dose on plasmid DNA expression after delivery in arginine/DNA complexes . Various amounts of plasmid DNA complexed with arginine peptide at an N/P ratio of 3:1 were intraperitoneally administered to mice, and mRNA levels were evaluated using real time RT-PCR. Total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Article Snippet: The thermal cycler protocol was set as follows: pre-incubation at 95°C for 10 s, amplification at 40 cycles at 95°C for 5 s, and 60°C for 40 s. For the mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA measurements, each sample was prepared following the manufacturer's instructions with a GAPDH primer set (Qiagen).

    Techniques: Plasmid Preparation, Expressing, Mouse Assay, Quantitative RT-PCR, Polymerase Chain Reaction, Amplification, Luciferase

    Duration of plasmid DNA expression . Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice, and total RNA was extracted from the organs at the indicated time points. The RNA extracts were transformed to cDNA using RT-PCR to serve as templates for nested PCR analysis. PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. The nested PCR products were separated on a 1.2% agarose gel.

    Journal: Genetic Vaccines and Therapy

    Article Title: Biodistribution and blood clearance of plasmid DNA administered in arginine peptide complexes

    doi: 10.1186/1479-0556-9-13

    Figure Lengend Snippet: Duration of plasmid DNA expression . Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice, and total RNA was extracted from the organs at the indicated time points. The RNA extracts were transformed to cDNA using RT-PCR to serve as templates for nested PCR analysis. PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. The nested PCR products were separated on a 1.2% agarose gel.

    Article Snippet: The thermal cycler protocol was set as follows: pre-incubation at 95°C for 10 s, amplification at 40 cycles at 95°C for 5 s, and 60°C for 40 s. For the mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA measurements, each sample was prepared following the manufacturer's instructions with a GAPDH primer set (Qiagen).

    Techniques: Plasmid Preparation, Expressing, Mouse Assay, Transformation Assay, Reverse Transcription Polymerase Chain Reaction, Nested PCR, Polymerase Chain Reaction, Amplification, Luciferase, Agarose Gel Electrophoresis