rt² qpcr primer assay  (Qiagen)


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    Name:
    RT² qPCR Primer Assay
    Description:
    RT² qPCR Primer Assays are specifically designed and experimentally verified for real time PCR analysis The rigorous assay verification criteria ensure PCR specificity and efficiency for reliable and accurate gene expression analysis results
    Catalog Number:
    330001
    Price:
    144
    Category:
    Assay PCR qPCR
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    Structured Review

    Qiagen rt² qpcr primer assay
    RT² qPCR Primer Assay
    RT² qPCR Primer Assays are specifically designed and experimentally verified for real time PCR analysis The rigorous assay verification criteria ensure PCR specificity and efficiency for reliable and accurate gene expression analysis results
    https://www.bioz.com/result/rt² qpcr primer assay/product/Qiagen
    Average 90 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    rt² qpcr primer assay - by Bioz Stars, 2020-01
    90/100 stars

    Images

    1) Product Images from "Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats"

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00380.2014

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,
    Figure Legend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Techniques Used: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric
    Figure Legend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Techniques Used: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P
    Figure Legend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Techniques Used: Expressing

    2) Product Images from "Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats"

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00380.2014

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,
    Figure Legend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Techniques Used: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric
    Figure Legend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Techniques Used: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P
    Figure Legend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Techniques Used: Expressing

    3) Product Images from "Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats"

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00380.2014

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,
    Figure Legend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Techniques Used: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric
    Figure Legend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Techniques Used: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P
    Figure Legend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Techniques Used: Expressing

    4) Product Images from "Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells"

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0173035

    Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P
    Figure Legend Snippet: Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P

    Techniques Used: Expressing, Western Blot

    Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.
    Figure Legend Snippet: Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.

    Techniques Used: Activation Assay

    Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P
    Figure Legend Snippet: Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P

    Techniques Used: Expressing, Binding Assay, Western Blot

    5) Product Images from "Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells"

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0173035

    Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P
    Figure Legend Snippet: Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P

    Techniques Used: Expressing, Western Blot

    Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.
    Figure Legend Snippet: Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.

    Techniques Used: Activation Assay

    Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P
    Figure Legend Snippet: Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P

    Techniques Used: Expressing, Binding Assay, Western Blot

    6) Product Images from "Effects of X-chromosome Tenomodulin Genetic Variants on Obesity in a Children’s Cohort and Implications of the Gene in Adipocyte Metabolism"

    Article Title: Effects of X-chromosome Tenomodulin Genetic Variants on Obesity in a Children’s Cohort and Implications of the Gene in Adipocyte Metabolism

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-40482-0

    TNMD promotes adipogenesis and impairs lipid metabolism in human adipocytes. Human adipocytes were transfected with an adenovirus-5 containing a shRNA-TNMD and shRNA-control (scrambled) at day 14 of adipogenesis induction. (a–c ) Peroxisome proliferator-activated receptor gamma (PPARG), CCAAT/enhancer-binding protein alpha (CEBPA) and angiopoietin-like 4 (ANGPTL4) mRNA and protein levels were determined in the shRNA-TNMD and shRNA-control adipocytes. Protein levels in cell lysates were analyzed by Western blotting using specific antibodies against PPARG, CEBPA and ANGPTL4, normalized to the internal control (α-tubulin), and expressed as fold-change; the lower section shows a representative crop blot. (d – f) Hormone-sensitive lipase ( HSL ) gene expression, adipose triglyceride lipase ( ATGL ), and perilipin ( PLIN ) gene expression were determined in the shRNA-TNMD and shRNA-control adipocytes. ( g ) Glycerol levels (µM) in cell supernatants after treatment with shRNA-TNMD. All values are expressed as the means ± SEM of three independent experiments. Significant differences were identified using the nonparametric Mann-Whitney U test; *P-value
    Figure Legend Snippet: TNMD promotes adipogenesis and impairs lipid metabolism in human adipocytes. Human adipocytes were transfected with an adenovirus-5 containing a shRNA-TNMD and shRNA-control (scrambled) at day 14 of adipogenesis induction. (a–c ) Peroxisome proliferator-activated receptor gamma (PPARG), CCAAT/enhancer-binding protein alpha (CEBPA) and angiopoietin-like 4 (ANGPTL4) mRNA and protein levels were determined in the shRNA-TNMD and shRNA-control adipocytes. Protein levels in cell lysates were analyzed by Western blotting using specific antibodies against PPARG, CEBPA and ANGPTL4, normalized to the internal control (α-tubulin), and expressed as fold-change; the lower section shows a representative crop blot. (d – f) Hormone-sensitive lipase ( HSL ) gene expression, adipose triglyceride lipase ( ATGL ), and perilipin ( PLIN ) gene expression were determined in the shRNA-TNMD and shRNA-control adipocytes. ( g ) Glycerol levels (µM) in cell supernatants after treatment with shRNA-TNMD. All values are expressed as the means ± SEM of three independent experiments. Significant differences were identified using the nonparametric Mann-Whitney U test; *P-value

    Techniques Used: Transfection, shRNA, Binding Assay, Western Blot, Expressing, MANN-WHITNEY

    7) Product Images from "PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro"

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174968

    Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used:

    Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used: Activity Assay, Software

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    Amplification:

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    Article Snippet: .. A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer. .. PCR cycle conditions were 95°C for 10 sec and 50 cycles of 9°C for 20 sec and 60°C for 30 sec.

    Article Title: CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target
    Article Snippet: .. CAIX and 18S specific primers (CAIX: Cat. No. PPH01751A; 18S: Cat. No. 330001 PPH05666E, Qiagen, Hilden, Germany) were used for amplification of cDNA, which was detected with Maxima SYBR Green (Thermo Fisher Scientific Inc., Waltham, MA, USA) in a Lightcycler 4800 (Roche, Penzberg, Germany). .. Patient tissue was analyzed in duplicates and triplicates, cell culture data in duplicates, in vivo data included tumour RNA of all animals of each treatment group (n = 9/10, respectively).

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    Article Snippet: cDNA preparation and validation RT-PCR Validation for the gene expression of Lipocalin 2 was carried out on individual cDNA samples from all the groups using the RT² qPCR Primer Assay (Qiagen). .. The Real-time qPCR (RT-PCR) were run in 96-well plates using the standard amplification conditions described for the 7500 RT-PCR system and 40 ng cDNA, 10 μl 2× Power SYBR green master mix (Applied Biosystems, Foster City, CA), and 1 μl of gene-specific oligonucleotide primers.

    Synthesized:

    Article Title: Plag1 and Plagl2 have overlapping and distinct functions in telencephalic development
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    Quantitative RT-PCR:

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    SYBR Green Assay:

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    Article Snippet: .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen). .. To evaluate the secretion of lysosomal enzymes into the culture medium, the cells were washed with PBS and incubated with fresh culture medium approximately 16 h before the analysis.

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats
    Article Snippet: The following materials were used: atropine sulfate, carbachol, guanethidine, TTX, devazepide, cholestyramine, deoxycholic acid, cholic acid, oleanolic acid, GW4064, SQ 22,536, BAPTA (Sigma-Aldrich, St. Louis, MO); N G -nitro- l -arginine methyl ester ( l -NAME, Research Biochemicals International, Natick, MA); Rp-cAMP (Santa Cruz Biotechnology, Dallas, TX); TRIzol [Life Sciences Solutions (Thermo Fisher Scientific), Carlsbad, CA]. iScript cDNA Synthesis Kit and iQ SYBR Green Supermix were purchased from Bio-Rad Laboratories (Philadelphia, PA). .. RT-qPCR primer assays for GAPDH (330001 PPR06557A), NOS1 (330001 PPR44930E), and GPBAR1 (TGR5, 330001 PPR49442A) were purchased from QIAGEN (Valencia, CA).

    Article Title: Defining the Roles of IFN-γ and IL-17A in Inflammation and Protection against Helicobacter pylori Infection
    Article Snippet: cDNA preparation and validation RT-PCR Validation for the gene expression of Lipocalin 2 was carried out on individual cDNA samples from all the groups using the RT² qPCR Primer Assay (Qiagen). .. The Real-time qPCR (RT-PCR) were run in 96-well plates using the standard amplification conditions described for the 7500 RT-PCR system and 40 ng cDNA, 10 μl 2× Power SYBR green master mix (Applied Biosystems, Foster City, CA), and 1 μl of gene-specific oligonucleotide primers.

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: .. RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen). ..

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Template quantification was performed with 7500 fast real-time PCR system (4351106, Applied Biosystems, Forster City, USA) using power fast SYBR Green Master Mix (4385612, Applied Biosystems, Forster City, USA) as dye layer and the relative standard curve calculation method. .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Incubation:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen). .. To evaluate the secretion of lysosomal enzymes into the culture medium, the cells were washed with PBS and incubated with fresh culture medium approximately 16 h before the analysis.

    Expressing:

    Article Title: Dynamic gene expression response to altered gravity in human T cells
    Article Snippet: Real-time PCR reactions were performed in triplicates using RT² SYBR® Green qPCR Mastermix (Qiagen) and RT² qPCR Primer Assay (Qiagen) on a CFX384 Real-Time PCR System (Bio-Rad, Germany), following manufacturer’s instructions. .. The data analysis using the comparative CT (delta delta CT) method was used for calculating relative quantitation of gene expression with the GAPDH housekeeping gene.

    Article Title: IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis
    Article Snippet: Quantitative RT-PCR was performed, as previously described, using primers for mouse IL-33, IL-4, IL-5, IL-13, CCL11 (forward, 5′-TGTCTCCCTCCACCATGCA-3′; reverse, 5′-GATCTTCTTACTGGTCATGATAAAGCA-3′), CCL24 (forward, 5′-TGCATCTCCCCATAGATTCTGT-3′; reverse, 5′-ACTCGGTTTTCTGGAATTTTCTTG-3′), and β-actin (forward, 5′-CAGGGTGTGATGGGAATG-3′; reverse, 5′-GTAGAAGGTGTGGTGCCAGAT-3′), and primers for human IL-33, IL-5 (PPH00692A-200), CCL11 (PPH0057B-200), CCL24 (PPH01162B-200), CCL26 (PPH0163E-200), and β-actin (330001 PPH00073E-200) (all from Qiagen), and on an Applied Biosystems Step Plus machine (Applied Biosystems). .. Expression of all target mRNA transcripts was normalized to β-actin, and reported as relative fold difference among groups within an experiment, with control group set arbitrarily at 1.

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: Paragraph title: Supporting information Protocol scheme for phagocytosis quantification. Spectral imaging microscopy of fluorescence in RAW 264.7 macrophages and CL1 tumor cell co-cultures. PEDF expression stimulates the migration of RAW 264.7 cells towards 2D conventional prostate tumor cell mono-culture in vitro . Production of Superoxide Radicals in RAW 264.7 cells using the WST-1 Tetrazolium-based in vitro assay. Phagocytosis visualization in PCa/RAW 264.7 co-cultures treated with PNPLA2 and ATP5B inhibitors. mRNAs expression levels of Angiostatin receptors in RAW 264.7 cells. ... Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Article Title: Plag1 and Plagl2 have overlapping and distinct functions in telencephalic development
    Article Snippet: RNA was extracted with TRIzol reagent following the instructions of the manufacturer (Thermo Fisher Scientific, Cat#15596026). cDNA was synthesized and RT-qPCR was performed using a RT2 primer assay kit (Qiagen, 330001) according to the manufacturer's instructions. .. We used the delta-delta Ct method to calculate relative expression levels, using three housekeeping genes to normalize ( Gapdh , B2M , Hrpt ).

    Article Title: Defining the Roles of IFN-γ and IL-17A in Inflammation and Protection against Helicobacter pylori Infection
    Article Snippet: .. cDNA preparation and validation RT-PCR Validation for the gene expression of Lipocalin 2 was carried out on individual cDNA samples from all the groups using the RT² qPCR Primer Assay (Qiagen). .. The Real-time qPCR (RT-PCR) were run in 96-well plates using the standard amplification conditions described for the 7500 RT-PCR system and 40 ng cDNA, 10 μl 2× Power SYBR green master mix (Applied Biosystems, Foster City, CA), and 1 μl of gene-specific oligonucleotide primers.

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen). .. The relative gene expression of il-2 has been determined by comparative cycle threshold analysis and the data were normalized to the internal housekeeping gene 18s rRNA and the relative mRNA level in the untreated group (untreated PHA-L-activated).

    Article Title: Structural and Functional Rescue of Chronic Metabolically Stressed Optic Nerves through Respiration
    Article Snippet: Hprt was used as the housekeeping gene, chosen after a comparison of Actb , Rpl , Hprt , and GlucB showed that Hprt had the most stable gene expression across age and strain using optic nerve mRNA. .. Sod2 mRNA levels were analyzed by using the PAMM-087Z RT2 Profiler PCR Array for Mouse Mitochondria (catalog #330001, QIAGEN) run on an Applied Biosystems QuantStudio 6K Flex 384 instrument.

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model
    Article Snippet: .. Commercially available pre-validated primers (Rt² qPCR Primer Assay Qiagen #330001 series) were employed to assess the expression of Tlr4 (#PPR45931B; NM_0191178), Nlrp3 (#PPR56639A; NM_001191642), Casp1 (#PPR06427A; NM_012762), Il1b (#PPR06480B; NM_031512) and the housekeeping gene Actb (#PPR06570C; NM_031144). .. Both PCR-array and qPCR analysis were performed using a ViiA™7 Real-Time PCR System (Life Technologies do Brasil, São Paulo) and the results assessed with the 12 K Flex QuantStudioTM software (Applied Biosystems, Foster City, CA).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: HMGB1 Promotes Mitochondrial Dysfunction–Triggered Striatal Neurodegeneration via Autophagy and Apoptosis Activation
    Article Snippet: The sequences of forward and reverse oligonucleotide primers used for amplification of specific gene sequences of HMGB-1 (forward primer: 5′-ATG GGC AAA GGA GAT CCT A-3′ ; reverse primer: 5′-ATT CAT CAT CAT CAT CTT CT-3′ ) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH ; forward primer: 5′-TCC CTC AAG ATT GTC AGC AA-3′ ; reverse primer: 5′-AGA TCC ACA ACG GAT ACA TT-3′ ). .. A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer.

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The following primers were used: LERP1-forward: 5′ TAA TAC GAC TCA CTA TAG GCC TGC AGG TGA CAA AAT GCG 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GCT GCA ACT ATT GGA TTG TAG ACC CTC 3′, LERP2-forward: 5′ TAA TAC GAC TCA CTA TAG GCA GCT CGC ACT TTG CTT AAG GAT G 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GCG TTG AGA GCT CCG AGG TGT TG 3′ and Rho1 (control dsRNA) forward: 5′ TAA TAC GAC TCA CTA TAG GTT TGT TTT GTG TTT AGT TCG GC 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GAT CAA GAA CAA CCA GAA CAT CG 3′. .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Article Title: Plag1 and Plagl2 have overlapping and distinct functions in telencephalic development
    Article Snippet: .. RNA was extracted with TRIzol reagent following the instructions of the manufacturer (Thermo Fisher Scientific, Cat15596026). cDNA was synthesized and RT-qPCR was performed using a RT2 primer assay kit (Qiagen, 330001) according to the manufacturer's instructions. .. The following RT2 qPCR primers were obtained from Qiagen: Gapdh (PPM02946E), B2m (PPM03562A), Hrpt (PPM03559F), Plag1 (PPM30678A), Plagl2 (PPM30603B) and Zac1 (PPM03537F). qPCR was performed with cortices from three embryos of each genotype and with three technical replicates for each biological replicate.

    Derivative Assay:

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model
    Article Snippet: For this, cDNA obtained from 9 animals of each experimental group was distributed among three pools, derived from 3 animals each. .. Commercially available pre-validated primers (Rt² qPCR Primer Assay Qiagen #330001 series) were employed to assess the expression of Tlr4 (#PPR45931B; NM_0191178), Nlrp3 (#PPR56639A; NM_001191642), Casp1 (#PPR06427A; NM_012762), Il1b (#PPR06480B; NM_031512) and the housekeeping gene Actb (#PPR06570C; NM_031144).

    Countercurrent Chromatography:

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Specific primer pairs were designed using primer 3 software ( http://frodo.wi.mit.edu ): L19 sense, 5’- GCG GAA GGG TAC AGC CAA T-3’ , L19-R antisense, 5’- GCA GCC GGC GCA AA-3’ ; decidual PRL sense 5’-AAG CTG TAG AGA TTG AGG AGC AAA C-3’ and decidual PRL antisense, 5’-TCA GGA TGA ACC TGG CTG ACT A-3’ ; IGFBP1 sense, 5’-CGA AGG CTC TCC ATG TCA CCA-3’ and IGFBP1 antisense, 5’-TGT CTC CTG TGC CTT GGC TAA AC-3’ ; 11βHSD1 sense, 5’-AGC AAG TTT GCT TTG GAT GG-3’ and 11βHSD1 antisense, 5’-AGA GCT CCC CCT TTG ATG AT-3’ ; CRABP2 sense, 5’-TGT GAG CAG AAG CTC CTG AAG-3’ and CRABP2 antisense 5’-GTT CTA CCT GTG GCC ACT CAC T-3’ ; RARα sense, 5’- GCC CAG CTC ACC ACA TCT TC-3’ and RARα antisense, 5’- GGA GCA ATG GCT TGT GAG TTC T-3’ ; RXRα sense, 5’- GTC CTT GGA GGC CTA CTG CAA-3’ and RXRα antisense, 5’- CCG ATG AGC TTG AAG AAG AAG AGA T-3’ ; PPARβ/δ sense, 5’- ACT GAC CCA ACT GAT CCT GCT C-3’ and PPARβ/δ antisense, 5’- GCC TGG CAA ACC AGT GTG AA-3’ ; DHRS3 sense, 5’-GAA GCT GTG CAG CTC AAC CA-3’ and DHRS3 antisense, 5’-CAT GCA GGT GTA GGT TCC TGA GA-3’ ; RDH12 sense, 5’- GGG AGT CTG CTG CCA GTG AA-3’ and RDH12 antisense, 5’- CCA TCA GCT GTC TTG GAA TAT GGA-3’ ; RBP4 sense, 5’- AGG CTC GCT TCT CTG GGA CC-3’ and RBP4 antisense, 5’- CCC TTG GCT GTG GCG CTC AT-3’ . .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Transfection:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: Mock-treated and mock-depleted cells were transfected without the addition of dsRNA or with Rho1 dsRNA, respectively. .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Pulse Chase:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen). .. Pulse-chase labeling experiments were performed with S2 cells that were treated with LERP RNAi for 5 days or mock-treated, as described in with minor modifications.

    Protease Inhibitor:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen). .. The S2 cells were lysed in 1% Triton X-100/PBS containing a protease inhibitor cocktail (Complete, Roche) and the activities of β-hexosaminidase, β-glucuronidase, α-mannosidase, β-mannosidase and β-galactosidase were determined as described below.

    Cell Culture:

    Article Title: CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target
    Article Snippet: CAIX and 18S specific primers (CAIX: Cat. No. PPH01751A; 18S: Cat. No. 330001 PPH05666E, Qiagen, Hilden, Germany) were used for amplification of cDNA, which was detected with Maxima SYBR Green (Thermo Fisher Scientific Inc., Waltham, MA, USA) in a Lightcycler 4800 (Roche, Penzberg, Germany). .. Patient tissue was analyzed in duplicates and triplicates, cell culture data in duplicates, in vivo data included tumour RNA of all animals of each treatment group (n = 9/10, respectively).

    Generated:

    Article Title: HMGB1 Promotes Mitochondrial Dysfunction–Triggered Striatal Neurodegeneration via Autophagy and Apoptosis Activation
    Article Snippet: Total RNA was extracted from the dissected striata with the RNAiso Reagent kit (Takara, DaLian, China). cDNA was generated by reverse-transcription of 2 μg total RNA using random primers and Primescript RT Reagent Kit (Takara) in a total reaction volume of 20 μL according to the manufacturer’s instructions. .. A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer.

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: And complementary DNA was generated using the SuperScript II Reverse Transcriptase for RT-PCR kit (18064, Invitrogen Ltd. Life technology, California, USA). .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Polymerase Chain Reaction:

    Article Title: HMGB1 Promotes Mitochondrial Dysfunction–Triggered Striatal Neurodegeneration via Autophagy and Apoptosis Activation
    Article Snippet: A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer. .. PCR cycle conditions were 95°C for 10 sec and 50 cycles of 9°C for 20 sec and 60°C for 30 sec.

    Article Title: Dynamic gene expression response to altered gravity in human T cells
    Article Snippet: Real-time PCR reactions were performed in triplicates using RT² SYBR® Green qPCR Mastermix (Qiagen) and RT² qPCR Primer Assay (Qiagen) on a CFX384 Real-Time PCR System (Bio-Rad, Germany), following manufacturer’s instructions. .. Dissociation curve analyses were carried out at the end of each run for PCR product verification.

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: PCR was performed with gene-specific primers flanked by the T7 RNA polymerase promoter sequence at the 5′-ends, as described in . .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Article Title: Structural and Functional Rescue of Chronic Metabolically Stressed Optic Nerves through Respiration
    Article Snippet: .. Sod2 mRNA levels were analyzed by using the PAMM-087Z RT2 Profiler PCR Array for Mouse Mitochondria (catalog #330001, QIAGEN) run on an Applied Biosystems QuantStudio 6K Flex 384 instrument. .. Fresh optic nerve tissue from D2 and D2G ONs was flash frozen in liquid nitrogen until homogenization in T-PER buffer (Thermo Fisher Scientific) with HALT protease and phosphatase inhibitors (catalog #78442, Thermo Fisher Scientific) using three 3 s pulses at 10% amplitude from a Branson Sonicator.

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model
    Article Snippet: The cDNAs were mixed with the appropriate PCR master mix buffer (Rt² Syber Green ROX qPCR Primer Assay QiagenN® #330523) and analyzed on specific array plates (Rat Innate & Adaptive Immune Responses Rt² Profiler™ PCR, Qiagen®, #330231-PARN052Zc). .. Commercially available pre-validated primers (Rt² qPCR Primer Assay Qiagen #330001 series) were employed to assess the expression of Tlr4 (#PPR45931B; NM_0191178), Nlrp3 (#PPR56639A; NM_001191642), Casp1 (#PPR06427A; NM_012762), Il1b (#PPR06480B; NM_031512) and the housekeeping gene Actb (#PPR06570C; NM_031144).

    Imaging:

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: Paragraph title: Supporting information Protocol scheme for phagocytosis quantification. Spectral imaging microscopy of fluorescence in RAW 264.7 macrophages and CL1 tumor cell co-cultures. PEDF expression stimulates the migration of RAW 264.7 cells towards 2D conventional prostate tumor cell mono-culture in vitro . Production of Superoxide Radicals in RAW 264.7 cells using the WST-1 Tetrazolium-based in vitro assay. Phagocytosis visualization in PCa/RAW 264.7 co-cultures treated with PNPLA2 and ATP5B inhibitors. mRNAs expression levels of Angiostatin receptors in RAW 264.7 cells. ... Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Sequencing:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: PCR was performed with gene-specific primers flanked by the T7 RNA polymerase promoter sequence at the 5′-ends, as described in . .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Cellular Antioxidant Activity Assay:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The following primers were used: LERP1-forward: 5′ TAA TAC GAC TCA CTA TAG GCC TGC AGG TGA CAA AAT GCG 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GCT GCA ACT ATT GGA TTG TAG ACC CTC 3′, LERP2-forward: 5′ TAA TAC GAC TCA CTA TAG GCA GCT CGC ACT TTG CTT AAG GAT G 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GCG TTG AGA GCT CCG AGG TGT TG 3′ and Rho1 (control dsRNA) forward: 5′ TAA TAC GAC TCA CTA TAG GTT TGT TTT GTG TTT AGT TCG GC 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GAT CAA GAA CAA CCA GAA CAT CG 3′. .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Specific primer pairs were designed using primer 3 software ( http://frodo.wi.mit.edu ): L19 sense, 5’- GCG GAA GGG TAC AGC CAA T-3’ , L19-R antisense, 5’- GCA GCC GGC GCA AA-3’ ; decidual PRL sense 5’-AAG CTG TAG AGA TTG AGG AGC AAA C-3’ and decidual PRL antisense, 5’-TCA GGA TGA ACC TGG CTG ACT A-3’ ; IGFBP1 sense, 5’-CGA AGG CTC TCC ATG TCA CCA-3’ and IGFBP1 antisense, 5’-TGT CTC CTG TGC CTT GGC TAA AC-3’ ; 11βHSD1 sense, 5’-AGC AAG TTT GCT TTG GAT GG-3’ and 11βHSD1 antisense, 5’-AGA GCT CCC CCT TTG ATG AT-3’ ; CRABP2 sense, 5’-TGT GAG CAG AAG CTC CTG AAG-3’ and CRABP2 antisense 5’-GTT CTA CCT GTG GCC ACT CAC T-3’ ; RARα sense, 5’- GCC CAG CTC ACC ACA TCT TC-3’ and RARα antisense, 5’- GGA GCA ATG GCT TGT GAG TTC T-3’ ; RXRα sense, 5’- GTC CTT GGA GGC CTA CTG CAA-3’ and RXRα antisense, 5’- CCG ATG AGC TTG AAG AAG AAG AGA T-3’ ; PPARβ/δ sense, 5’- ACT GAC CCA ACT GAT CCT GCT C-3’ and PPARβ/δ antisense, 5’- GCC TGG CAA ACC AGT GTG AA-3’ ; DHRS3 sense, 5’-GAA GCT GTG CAG CTC AAC CA-3’ and DHRS3 antisense, 5’-CAT GCA GGT GTA GGT TCC TGA GA-3’ ; RDH12 sense, 5’- GGG AGT CTG CTG CCA GTG AA-3’ and RDH12 antisense, 5’- CCA TCA GCT GTC TTG GAA TAT GGA-3’ ; RBP4 sense, 5’- AGG CTC GCT TCT CTG GGA CC-3’ and RBP4 antisense, 5’- CCC TTG GCT GTG GCG CTC AT-3’ . .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    In Vivo:

    Article Title: CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target
    Article Snippet: CAIX and 18S specific primers (CAIX: Cat. No. PPH01751A; 18S: Cat. No. 330001 PPH05666E, Qiagen, Hilden, Germany) were used for amplification of cDNA, which was detected with Maxima SYBR Green (Thermo Fisher Scientific Inc., Waltham, MA, USA) in a Lightcycler 4800 (Roche, Penzberg, Germany). .. Patient tissue was analyzed in duplicates and triplicates, cell culture data in duplicates, in vivo data included tumour RNA of all animals of each treatment group (n = 9/10, respectively).

    Fluorescence:

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: Paragraph title: Supporting information Protocol scheme for phagocytosis quantification. Spectral imaging microscopy of fluorescence in RAW 264.7 macrophages and CL1 tumor cell co-cultures. PEDF expression stimulates the migration of RAW 264.7 cells towards 2D conventional prostate tumor cell mono-culture in vitro . Production of Superoxide Radicals in RAW 264.7 cells using the WST-1 Tetrazolium-based in vitro assay. Phagocytosis visualization in PCa/RAW 264.7 co-cultures treated with PNPLA2 and ATP5B inhibitors. mRNAs expression levels of Angiostatin receptors in RAW 264.7 cells. ... Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Isolation:

    Article Title: IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis
    Article Snippet: Total RNA was isolated from patient endoscopic biopsy specimens and mouse ilea using the RNeasy Mini Kit (Qiagen, Germantown, MD) and reverse transcribed (RNA-to-cDNA kit; Applied Biosystems, Forest City, CA), both according to the manufacturer's instructions. .. Quantitative RT-PCR was performed, as previously described, using primers for mouse IL-33, IL-4, IL-5, IL-13, CCL11 (forward, 5′-TGTCTCCCTCCACCATGCA-3′; reverse, 5′-GATCTTCTTACTGGTCATGATAAAGCA-3′), CCL24 (forward, 5′-TGCATCTCCCCATAGATTCTGT-3′; reverse, 5′-ACTCGGTTTTCTGGAATTTTCTTG-3′), and β-actin (forward, 5′-CAGGGTGTGATGGGAATG-3′; reverse, 5′-GTAGAAGGTGTGGTGCCAGAT-3′), and primers for human IL-33, IL-5 (PPH00692A-200), CCL11 (PPH0057B-200), CCL24 (PPH01162B-200), CCL26 (PPH0163E-200), and β-actin (330001 PPH00073E-200) (all from Qiagen), and on an Applied Biosystems Step Plus machine (Applied Biosystems).

    Article Title: CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target
    Article Snippet: Reverse transcription quantitative PCR (RT qPCR) Total RNA was isolated with RNeasy Mini Kit (Qiagen, Hilden, Germany) in accordance to the manufacturer´s protocol and reversely transcribed with Quantitect Reverse Transcription Kit (Qiagen, Hilden, Germany). .. CAIX and 18S specific primers (CAIX: Cat. No. PPH01751A; 18S: Cat. No. 330001 PPH05666E, Qiagen, Hilden, Germany) were used for amplification of cDNA, which was detected with Maxima SYBR Green (Thermo Fisher Scientific Inc., Waltham, MA, USA) in a Lightcycler 4800 (Roche, Penzberg, Germany).

    Article Title: Structural and Functional Rescue of Chronic Metabolically Stressed Optic Nerves through Respiration
    Article Snippet: Fresh optic nerve tissue was collected from D2 and D2G ONs, and mRNA was isolated using TRIzol extraction (Thermo Fisher Scientific). mRNA was converted to cDNA (Verso cDNA Synthesis Kit, catalog #AB1453B, Thermo Fisher Scientific) then analyzed using real-time quantitative PCR with an ABI 7900HT instrument (Applied Biosystems) and TaqMan Gene Expression Assays with a FAM reporter dye at the 5′ end of the TaqMan MGB probe. .. Sod2 mRNA levels were analyzed by using the PAMM-087Z RT2 Profiler PCR Array for Mouse Mitochondria (catalog #330001, QIAGEN) run on an Applied Biosystems QuantStudio 6K Flex 384 instrument.

    Size-exclusion Chromatography:

    Article Title: HMGB1 Promotes Mitochondrial Dysfunction–Triggered Striatal Neurodegeneration via Autophagy and Apoptosis Activation
    Article Snippet: A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer. .. PCR cycle conditions were 95°C for 10 sec and 50 cycles of 9°C for 20 sec and 60°C for 30 sec.

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen). .. The real-time thermal cycler program comprised an initial denaturation step at 95° C for 10 min followed by a two-step program with 40 cycles (95°C, 15 sec; and 60°C, 60 sec).

    Labeling:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen). .. Pulse-chase labeling experiments were performed with S2 cells that were treated with LERP RNAi for 5 days or mock-treated, as described in with minor modifications.

    Purification:

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: The quantity and purity of extracted RNA was measured by nanodrop spectrophotometry (Peqlab, Erlangen, Gemany) and purified RNA was reverse transcribed using the RT2 First Strand Kit (Qiagen). .. RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Defining the Roles of IFN-γ and IL-17A in Inflammation and Protection against Helicobacter pylori Infection
    Article Snippet: .. cDNA preparation and validation RT-PCR Validation for the gene expression of Lipocalin 2 was carried out on individual cDNA samples from all the groups using the RT² qPCR Primer Assay (Qiagen). .. The Real-time qPCR (RT-PCR) were run in 96-well plates using the standard amplification conditions described for the 7500 RT-PCR system and 40 ng cDNA, 10 μl 2× Power SYBR green master mix (Applied Biosystems, Foster City, CA), and 1 μl of gene-specific oligonucleotide primers.

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: .. RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen). ..

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: And complementary DNA was generated using the SuperScript II Reverse Transcriptase for RT-PCR kit (18064, Invitrogen Ltd. Life technology, California, USA). .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Microscopy:

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: Paragraph title: Supporting information Protocol scheme for phagocytosis quantification. Spectral imaging microscopy of fluorescence in RAW 264.7 macrophages and CL1 tumor cell co-cultures. PEDF expression stimulates the migration of RAW 264.7 cells towards 2D conventional prostate tumor cell mono-culture in vitro . Production of Superoxide Radicals in RAW 264.7 cells using the WST-1 Tetrazolium-based in vitro assay. Phagocytosis visualization in PCa/RAW 264.7 co-cultures treated with PNPLA2 and ATP5B inhibitors. mRNAs expression levels of Angiostatin receptors in RAW 264.7 cells. ... Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Activated Clotting Time Assay:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: The following primers were used: LERP1-forward: 5′ TAA TAC GAC TCA CTA TAG GCC TGC AGG TGA CAA AAT GCG 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GCT GCA ACT ATT GGA TTG TAG ACC CTC 3′, LERP2-forward: 5′ TAA TAC GAC TCA CTA TAG GCA GCT CGC ACT TTG CTT AAG GAT G 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GCG TTG AGA GCT CCG AGG TGT TG 3′ and Rho1 (control dsRNA) forward: 5′ TAA TAC GAC TCA CTA TAG GTT TGT TTT GTG TTT AGT TCG GC 3′ and reverse: 5′ TAA TAC GAC TCA CTA TAG GAT CAA GAA CAA CCA GAA CAT CG 3′. .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Specific primer pairs were designed using primer 3 software ( http://frodo.wi.mit.edu ): L19 sense, 5’- GCG GAA GGG TAC AGC CAA T-3’ , L19-R antisense, 5’- GCA GCC GGC GCA AA-3’ ; decidual PRL sense 5’-AAG CTG TAG AGA TTG AGG AGC AAA C-3’ and decidual PRL antisense, 5’-TCA GGA TGA ACC TGG CTG ACT A-3’ ; IGFBP1 sense, 5’-CGA AGG CTC TCC ATG TCA CCA-3’ and IGFBP1 antisense, 5’-TGT CTC CTG TGC CTT GGC TAA AC-3’ ; 11βHSD1 sense, 5’-AGC AAG TTT GCT TTG GAT GG-3’ and 11βHSD1 antisense, 5’-AGA GCT CCC CCT TTG ATG AT-3’ ; CRABP2 sense, 5’-TGT GAG CAG AAG CTC CTG AAG-3’ and CRABP2 antisense 5’-GTT CTA CCT GTG GCC ACT CAC T-3’ ; RARα sense, 5’- GCC CAG CTC ACC ACA TCT TC-3’ and RARα antisense, 5’- GGA GCA ATG GCT TGT GAG TTC T-3’ ; RXRα sense, 5’- GTC CTT GGA GGC CTA CTG CAA-3’ and RXRα antisense, 5’- CCG ATG AGC TTG AAG AAG AAG AGA T-3’ ; PPARβ/δ sense, 5’- ACT GAC CCA ACT GAT CCT GCT C-3’ and PPARβ/δ antisense, 5’- GCC TGG CAA ACC AGT GTG AA-3’ ; DHRS3 sense, 5’-GAA GCT GTG CAG CTC AAC CA-3’ and DHRS3 antisense, 5’-CAT GCA GGT GTA GGT TCC TGA GA-3’ ; RDH12 sense, 5’- GGG AGT CTG CTG CCA GTG AA-3’ and RDH12 antisense, 5’- CCA TCA GCT GTC TTG GAA TAT GGA-3’ ; RBP4 sense, 5’- AGG CTC GCT TCT CTG GGA CC-3’ and RBP4 antisense, 5’- CCC TTG GCT GTG GCG CTC AT-3’ . .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Software:

    Article Title: HMGB1 Promotes Mitochondrial Dysfunction–Triggered Striatal Neurodegeneration via Autophagy and Apoptosis Activation
    Article Snippet: A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer. .. Threshold cycle values, Ct, which correlates inversely with the target mRNA levels, were calculated with the second derivative maximum algorithm provided by the iCycler software.

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model
    Article Snippet: Commercially available pre-validated primers (Rt² qPCR Primer Assay Qiagen #330001 series) were employed to assess the expression of Tlr4 (#PPR45931B; NM_0191178), Nlrp3 (#PPR56639A; NM_001191642), Casp1 (#PPR06427A; NM_012762), Il1b (#PPR06480B; NM_031512) and the housekeeping gene Actb (#PPR06570C; NM_031144). .. Both PCR-array and qPCR analysis were performed using a ViiA™7 Real-Time PCR System (Life Technologies do Brasil, São Paulo) and the results assessed with the 12 K Flex QuantStudioTM software (Applied Biosystems, Foster City, CA).

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Specific primer pairs were designed using primer 3 software ( http://frodo.wi.mit.edu ): L19 sense, 5’- GCG GAA GGG TAC AGC CAA T-3’ , L19-R antisense, 5’- GCA GCC GGC GCA AA-3’ ; decidual PRL sense 5’-AAG CTG TAG AGA TTG AGG AGC AAA C-3’ and decidual PRL antisense, 5’-TCA GGA TGA ACC TGG CTG ACT A-3’ ; IGFBP1 sense, 5’-CGA AGG CTC TCC ATG TCA CCA-3’ and IGFBP1 antisense, 5’-TGT CTC CTG TGC CTT GGC TAA AC-3’ ; 11βHSD1 sense, 5’-AGC AAG TTT GCT TTG GAT GG-3’ and 11βHSD1 antisense, 5’-AGA GCT CCC CCT TTG ATG AT-3’ ; CRABP2 sense, 5’-TGT GAG CAG AAG CTC CTG AAG-3’ and CRABP2 antisense 5’-GTT CTA CCT GTG GCC ACT CAC T-3’ ; RARα sense, 5’- GCC CAG CTC ACC ACA TCT TC-3’ and RARα antisense, 5’- GGA GCA ATG GCT TGT GAG TTC T-3’ ; RXRα sense, 5’- GTC CTT GGA GGC CTA CTG CAA-3’ and RXRα antisense, 5’- CCG ATG AGC TTG AAG AAG AAG AGA T-3’ ; PPARβ/δ sense, 5’- ACT GAC CCA ACT GAT CCT GCT C-3’ and PPARβ/δ antisense, 5’- GCC TGG CAA ACC AGT GTG AA-3’ ; DHRS3 sense, 5’-GAA GCT GTG CAG CTC AAC CA-3’ and DHRS3 antisense, 5’-CAT GCA GGT GTA GGT TCC TGA GA-3’ ; RDH12 sense, 5’- GGG AGT CTG CTG CCA GTG AA-3’ and RDH12 antisense, 5’- CCA TCA GCT GTC TTG GAA TAT GGA-3’ ; RBP4 sense, 5’- AGG CTC GCT TCT CTG GGA CC-3’ and RBP4 antisense, 5’- CCC TTG GCT GTG GCG CTC AT-3’ . .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: HMGB1 Promotes Mitochondrial Dysfunction–Triggered Striatal Neurodegeneration via Autophagy and Apoptosis Activation
    Article Snippet: .. A 20-fold dilution of each cDNA was amplified in a 20-μL volume using the RT² qPCR Primer Assay (QIAGEN Inc., Valencia, CA), with 200 nM final concentrations of each primer. .. PCR cycle conditions were 95°C for 10 sec and 50 cycles of 9°C for 20 sec and 60°C for 30 sec.

    Article Title: Dynamic gene expression response to altered gravity in human T cells
    Article Snippet: .. Real-time PCR reactions were performed in triplicates using RT² SYBR® Green qPCR Mastermix (Qiagen) and RT² qPCR Primer Assay (Qiagen) on a CFX384 Real-Time PCR System (Bio-Rad, Germany), following manufacturer’s instructions. .. Dissociation curve analyses were carried out at the end of each run for PCR product verification.

    Article Title: CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target
    Article Snippet: Paragraph title: Reverse transcription quantitative PCR (RT qPCR) ... CAIX and 18S specific primers (CAIX: Cat. No. PPH01751A; 18S: Cat. No. 330001 PPH05666E, Qiagen, Hilden, Germany) were used for amplification of cDNA, which was detected with Maxima SYBR Green (Thermo Fisher Scientific Inc., Waltham, MA, USA) in a Lightcycler 4800 (Roche, Penzberg, Germany).

    Article Title: Plag1 and Plagl2 have overlapping and distinct functions in telencephalic development
    Article Snippet: RNA was extracted with TRIzol reagent following the instructions of the manufacturer (Thermo Fisher Scientific, Cat#15596026). cDNA was synthesized and RT-qPCR was performed using a RT2 primer assay kit (Qiagen, 330001) according to the manufacturer's instructions. .. The following RT2 qPCR primers were obtained from Qiagen: Gapdh (PPM02946E), B2m (PPM03562A), Hrpt (PPM03559F), Plag1 (PPM30678A), Plagl2 (PPM30603B) and Zac1 (PPM03537F). qPCR was performed with cortices from three embryos of each genotype and with three technical replicates for each biological replicate.

    Article Title: Defining the Roles of IFN-γ and IL-17A in Inflammation and Protection against Helicobacter pylori Infection
    Article Snippet: .. cDNA preparation and validation RT-PCR Validation for the gene expression of Lipocalin 2 was carried out on individual cDNA samples from all the groups using the RT² qPCR Primer Assay (Qiagen). .. The Real-time qPCR (RT-PCR) were run in 96-well plates using the standard amplification conditions described for the 7500 RT-PCR system and 40 ng cDNA, 10 μl 2× Power SYBR green master mix (Applied Biosystems, Foster City, CA), and 1 μl of gene-specific oligonucleotide primers.

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: .. RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen). ..

    Article Title: Structural and Functional Rescue of Chronic Metabolically Stressed Optic Nerves through Respiration
    Article Snippet: Quantitative PCR analysis was repeated three to six times using biological replicates; individual runs used biological replicate template in quadruplicate. .. Sod2 mRNA levels were analyzed by using the PAMM-087Z RT2 Profiler PCR Array for Mouse Mitochondria (catalog #330001, QIAGEN) run on an Applied Biosystems QuantStudio 6K Flex 384 instrument.

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model
    Article Snippet: .. Commercially available pre-validated primers (Rt² qPCR Primer Assay Qiagen #330001 series) were employed to assess the expression of Tlr4 (#PPR45931B; NM_0191178), Nlrp3 (#PPR56639A; NM_001191642), Casp1 (#PPR06427A; NM_012762), Il1b (#PPR06480B; NM_031512) and the housekeeping gene Actb (#PPR06570C; NM_031144). .. Both PCR-array and qPCR analysis were performed using a ViiA™7 Real-Time PCR System (Life Technologies do Brasil, São Paulo) and the results assessed with the 12 K Flex QuantStudioTM software (Applied Biosystems, Foster City, CA).

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Paragraph title: Real-Time Quantitative PCR (RTQ-PCR) ... We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    RNA Extraction:

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: Paragraph title: Total RNA extraction, reverse transcription and real-time PCR ... RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen).

    In Vitro:

    Article Title: The lysosomal enzyme receptor protein (LERP) is not essential, but is implicated in lysosomal function in Drosophila melanogaster
    Article Snippet: In vitro transcription was performed with the MEGAscript RNAi kit (Ambion) as instructed by the manufacturer. .. The level of knockdown relative to GAPDH (primers Cat. #330001 PPD03944A, Qiagen) was determined by quantitative RT-PCR using SYBR green master mix (SA Biosciences) and 10 μM primers to LERP (Cat. #330001 PPD10274A, Qiagen).

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: Paragraph title: Supporting information Protocol scheme for phagocytosis quantification. Spectral imaging microscopy of fluorescence in RAW 264.7 macrophages and CL1 tumor cell co-cultures. PEDF expression stimulates the migration of RAW 264.7 cells towards 2D conventional prostate tumor cell mono-culture in vitro . Production of Superoxide Radicals in RAW 264.7 cells using the WST-1 Tetrazolium-based in vitro assay. Phagocytosis visualization in PCa/RAW 264.7 co-cultures treated with PNPLA2 and ATP5B inhibitors. mRNAs expression levels of Angiostatin receptors in RAW 264.7 cells. ... Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Quantitation Assay:

    Article Title: Dynamic gene expression response to altered gravity in human T cells
    Article Snippet: Real-time PCR reactions were performed in triplicates using RT² SYBR® Green qPCR Mastermix (Qiagen) and RT² qPCR Primer Assay (Qiagen) on a CFX384 Real-Time PCR System (Bio-Rad, Germany), following manufacturer’s instructions. .. The data analysis using the comparative CT (delta delta CT) method was used for calculating relative quantitation of gene expression with the GAPDH housekeeping gene.

    Spectrophotometry:

    Article Title: Cyclotides Suppress Human T-Lymphocyte Proliferation by an Interleukin 2-Dependent Mechanism
    Article Snippet: The quantity and purity of extracted RNA was measured by nanodrop spectrophotometry (Peqlab, Erlangen, Gemany) and purified RNA was reverse transcribed using the RT2 First Strand Kit (Qiagen). .. RT-PCR reactions were carried out on a MyiQ kit (BioRad, Munich, Germany) in a final volume of 25 µL using RT² qPCR Primer Assay (for il-2 ) and SYBR® Green qPCR master mix (both from Qiagen).

    Concentration Assay:

    Article Title: IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis
    Article Snippet: Quantitative RT-PCR was performed, as previously described, using primers for mouse IL-33, IL-4, IL-5, IL-13, CCL11 (forward, 5′-TGTCTCCCTCCACCATGCA-3′; reverse, 5′-GATCTTCTTACTGGTCATGATAAAGCA-3′), CCL24 (forward, 5′-TGCATCTCCCCATAGATTCTGT-3′; reverse, 5′-ACTCGGTTTTCTGGAATTTTCTTG-3′), and β-actin (forward, 5′-CAGGGTGTGATGGGAATG-3′; reverse, 5′-GTAGAAGGTGTGGTGCCAGAT-3′), and primers for human IL-33, IL-5 (PPH00692A-200), CCL11 (PPH0057B-200), CCL24 (PPH01162B-200), CCL26 (PPH0163E-200), and β-actin (330001 PPH00073E-200) (all from Qiagen), and on an Applied Biosystems Step Plus machine (Applied Biosystems). .. Reaction mixture consisted of 15% volume first-strand synthesis in 20 μL total volume that included Power SYBR Green core reagents (Applied Biosystems) and 500 nmol/L final concentration of primers.

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model
    Article Snippet: After evaluation of RNA purity and final concentration, cDNA was synthesized through reverse transcription (RT) using the Rt² Easy First Strand kit (Qiagen® #330401). .. Commercially available pre-validated primers (Rt² qPCR Primer Assay Qiagen #330001 series) were employed to assess the expression of Tlr4 (#PPR45931B; NM_0191178), Nlrp3 (#PPR56639A; NM_001191642), Casp1 (#PPR06427A; NM_012762), Il1b (#PPR06480B; NM_031512) and the housekeeping gene Actb (#PPR06570C; NM_031144).

    Migration:

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: Paragraph title: Supporting information Protocol scheme for phagocytosis quantification. Spectral imaging microscopy of fluorescence in RAW 264.7 macrophages and CL1 tumor cell co-cultures. PEDF expression stimulates the migration of RAW 264.7 cells towards 2D conventional prostate tumor cell mono-culture in vitro . Production of Superoxide Radicals in RAW 264.7 cells using the WST-1 Tetrazolium-based in vitro assay. Phagocytosis visualization in PCa/RAW 264.7 co-cultures treated with PNPLA2 and ATP5B inhibitors. mRNAs expression levels of Angiostatin receptors in RAW 264.7 cells. ... Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    CTG Assay:

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells
    Article Snippet: Specific primer pairs were designed using primer 3 software ( http://frodo.wi.mit.edu ): L19 sense, 5’- GCG GAA GGG TAC AGC CAA T-3’ , L19-R antisense, 5’- GCA GCC GGC GCA AA-3’ ; decidual PRL sense 5’-AAG CTG TAG AGA TTG AGG AGC AAA C-3’ and decidual PRL antisense, 5’-TCA GGA TGA ACC TGG CTG ACT A-3’ ; IGFBP1 sense, 5’-CGA AGG CTC TCC ATG TCA CCA-3’ and IGFBP1 antisense, 5’-TGT CTC CTG TGC CTT GGC TAA AC-3’ ; 11βHSD1 sense, 5’-AGC AAG TTT GCT TTG GAT GG-3’ and 11βHSD1 antisense, 5’-AGA GCT CCC CCT TTG ATG AT-3’ ; CRABP2 sense, 5’-TGT GAG CAG AAG CTC CTG AAG-3’ and CRABP2 antisense 5’-GTT CTA CCT GTG GCC ACT CAC T-3’ ; RARα sense, 5’- GCC CAG CTC ACC ACA TCT TC-3’ and RARα antisense, 5’- GGA GCA ATG GCT TGT GAG TTC T-3’ ; RXRα sense, 5’- GTC CTT GGA GGC CTA CTG CAA-3’ and RXRα antisense, 5’- CCG ATG AGC TTG AAG AAG AAG AGA T-3’ ; PPARβ/δ sense, 5’- ACT GAC CCA ACT GAT CCT GCT C-3’ and PPARβ/δ antisense, 5’- GCC TGG CAA ACC AGT GTG AA-3’ ; DHRS3 sense, 5’-GAA GCT GTG CAG CTC AAC CA-3’ and DHRS3 antisense, 5’-CAT GCA GGT GTA GGT TCC TGA GA-3’ ; RDH12 sense, 5’- GGG AGT CTG CTG CCA GTG AA-3’ and RDH12 antisense, 5’- CCA TCA GCT GTC TTG GAA TAT GGA-3’ ; RBP4 sense, 5’- AGG CTC GCT TCT CTG GGA CC-3’ and RBP4 antisense, 5’- CCC TTG GCT GTG GCG CTC AT-3’ . .. We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

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    Qiagen rt² qpcr primer assay
    Rt² Qpcr Primer Assay, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rt² qpcr primer assay - by Bioz Stars, 2020-01
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