anti dna polymerase ii  (Millipore)


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    Name:
    DNA Polymerase I
    Description:
    DNA polymerase I yields two fragments small and large upon protease digestion The large fragment Klenow fragment loses the 5 exonuclease activity that is present in the intact holoenzyme However it retains both the polymerase 5 3 activity and the 3 5 exonuclease activity of the native enzyme
    Catalog Number:
    d8276
    Price:
    None
    Applications:
    Suitable for:. DNA sequencing by the Sanger dideoxy method. Synthesis of the complementary strand of cDNA. Filling in 5'-overhangs in double stranded DNA to form blunt ends. Mutagenesis of DNA with second strand synthesis using oligonucleotides. Labeling DNA by the random primer method
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    Structured Review

    Millipore anti dna polymerase ii
    <t>DNA</t> methyltransferase 1 <t>(DNMT1)-mediated</t> upregulation of CLDN6 expression by SB431542. ( A , B ) Real-time polymerase chain reaction (RT-PCR) and immunoblot analysis of DNMT1 and CLDN6, and densitometric analysis of relative gene expression levels after normalization to loading controls GAPDH and β-actin are presented. ( C ) DNMT1 activity assays. ( D ) Methylation-specific PCR (MSP) analysis of CpG island of CLDN6 promoter using bisulfite-treated genomic DNA isolated from MCF-7 and SKBR-3 cells. ( E ) CpG island methylation within the CLDN6 promoter region was measured by bisulfite sequencing in SKBR-3 cells. “Me” stands for methylated, and “U” stands for unmethylated. ( F ) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay to detect the binding of DNMT1 to the promoter of CLDN6 (** p
    DNA polymerase I yields two fragments small and large upon protease digestion The large fragment Klenow fragment loses the 5 exonuclease activity that is present in the intact holoenzyme However it retains both the polymerase 5 3 activity and the 3 5 exonuclease activity of the native enzyme
    https://www.bioz.com/result/anti dna polymerase ii/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti dna polymerase ii - by Bioz Stars, 2020-04
    99/100 stars

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    Images

    1) Product Images from "SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells"

    Article Title: SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms18091863

    DNA methyltransferase 1 (DNMT1)-mediated upregulation of CLDN6 expression by SB431542. ( A , B ) Real-time polymerase chain reaction (RT-PCR) and immunoblot analysis of DNMT1 and CLDN6, and densitometric analysis of relative gene expression levels after normalization to loading controls GAPDH and β-actin are presented. ( C ) DNMT1 activity assays. ( D ) Methylation-specific PCR (MSP) analysis of CpG island of CLDN6 promoter using bisulfite-treated genomic DNA isolated from MCF-7 and SKBR-3 cells. ( E ) CpG island methylation within the CLDN6 promoter region was measured by bisulfite sequencing in SKBR-3 cells. “Me” stands for methylated, and “U” stands for unmethylated. ( F ) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay to detect the binding of DNMT1 to the promoter of CLDN6 (** p
    Figure Legend Snippet: DNA methyltransferase 1 (DNMT1)-mediated upregulation of CLDN6 expression by SB431542. ( A , B ) Real-time polymerase chain reaction (RT-PCR) and immunoblot analysis of DNMT1 and CLDN6, and densitometric analysis of relative gene expression levels after normalization to loading controls GAPDH and β-actin are presented. ( C ) DNMT1 activity assays. ( D ) Methylation-specific PCR (MSP) analysis of CpG island of CLDN6 promoter using bisulfite-treated genomic DNA isolated from MCF-7 and SKBR-3 cells. ( E ) CpG island methylation within the CLDN6 promoter region was measured by bisulfite sequencing in SKBR-3 cells. “Me” stands for methylated, and “U” stands for unmethylated. ( F ) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay to detect the binding of DNMT1 to the promoter of CLDN6 (** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Methylation, Polymerase Chain Reaction, Isolation, Methylation Sequencing, Chromatin Immunoprecipitation, Binding Assay

    Related Articles

    Clone Assay:

    Article Title: Contrasting Effects of W781V and W780V Mutations in Helix N of Herpes Simplex Virus 1 and Human Cytomegalovirus DNA Polymerases on Antiviral Drug Susceptibility
    Article Snippet: .. The HSV-1 and HCMV DNA polymerase genes were cloned into pCITE4a (EMD BioScience, San Diego, CA) using the NdeI/NcoI and NdeI/SacI restriction sites to generate pCITE-UL30 and pCITE-UL54, respectively. .. The W781V and W780V mutations were introduced into the UL30 (pCITE-UL30) and UL54 (pCITE-UL54) genes, respectively, using a QuikChange site-directed mutagenesis kit (Stratagene) to produce plasmids pCITE-W781V and pCITE-W780V, respectively.

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: .. The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902. .. The gene (TK1903) encoding the large subunit of PolD (DP2) without the intein was cloned by GeneArt into pET-21a (Novagen) to generate pET-TK1903.

    Amplification:

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: .. The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902. .. The gene (TK1903) encoding the large subunit of PolD (DP2) without the intein was cloned by GeneArt into pET-21a (Novagen) to generate pET-TK1903.

    Positive Control:

    Article Title: SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells
    Article Snippet: .. Antibodies included anti-DNMT1 (1:500, Cell Signaling Technology), anti-IgG (Millipore), and anti-DNA polymerase II (Millipore); anti-IgG was the negative control and anti-DNA polymerase II was the positive control. .. Transfection With Short Hairpin RNA Cells were transfected with short hairpin RNA (shRNA) by using Lipofectamine 2000 (Invitrogen), following the procedure recommended by the manufacturer. shRNA targeting of CLDN6 (5’-GTGCAAGGTGTACGACTCA-3’) and a negative control shRNA were purchased from GeneChem Co. Ltd.

    Electrophoresis:

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: After electrophoresis (25 min, 25 V), air-dried and neutralized slides were stained with 2 μg/ml propidium iodide. .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin.

    Enzymatic Assay:

    Article Title: Guanine α-carboxy nucleoside phosphonate (G-α-CNP) shows a different inhibitory kinetic profile against the DNA polymerases of human immunodeficiency virus (HIV) and herpes viruses
    Article Snippet: Paragraph title: 2.6. Enzyme assay with HCMV DNA polymerase ... To perform the HCMV DNA polymerase assay, 4 μl of the TnT® reaction product was added to a 46 μl mixture to obtain 25 mM Tris.HCl pH 8.0, 100 mM (NH4 )2 SO4 (Sigma), 0.5 mM dithiothreitol, 10 mM MgCl2 , 0.2 mg/ml bovine serum albumin, 5 % glycerol (Acros, Geel, Belgium), 150 ng per μl activated calf thymus DNA (from Amersham Biosciences, Piscataway, N.J.), 100 μM of each of the three unlabeled dNTPs (GE Healthcare), and 0.5 μM of the rate-limiting tritium-labeled dNTP, and serial dilutions of the α-CNP.

    Single Cell Gel Electrophoresis:

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: Average comet tail moment was scored for duplicate slides (50 cells/slide) in randomly selected fields, and automated calculations were performed using Comet Assay II software (Perceptive Instruments, Suffolk, UK). .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin.

    Expressing:

    Article Title: Contrasting Effects of W781V and W780V Mutations in Helix N of Herpes Simplex Virus 1 and Human Cytomegalovirus DNA Polymerases on Antiviral Drug Susceptibility
    Article Snippet: Paragraph title: Expression of viral DNA polymerase proteins. ... The HSV-1 and HCMV DNA polymerase genes were cloned into pCITE4a (EMD BioScience, San Diego, CA) using the NdeI/NcoI and NdeI/SacI restriction sites to generate pCITE-UL30 and pCITE-UL54, respectively.

    Article Title: Guanine α-carboxy nucleoside phosphonate (G-α-CNP) shows a different inhibitory kinetic profile against the DNA polymerases of human immunodeficiency virus (HIV) and herpes viruses
    Article Snippet: Protein expression was performed with the TnT® SP6 Quick Coupled Transcription/Translation System (Promega) ( ). .. To perform the HCMV DNA polymerase assay, 4 μl of the TnT® reaction product was added to a 46 μl mixture to obtain 25 mM Tris.HCl pH 8.0, 100 mM (NH4 )2 SO4 (Sigma), 0.5 mM dithiothreitol, 10 mM MgCl2 , 0.2 mg/ml bovine serum albumin, 5 % glycerol (Acros, Geel, Belgium), 150 ng per μl activated calf thymus DNA (from Amersham Biosciences, Piscataway, N.J.), 100 μM of each of the three unlabeled dNTPs (GE Healthcare), and 0.5 μM of the rate-limiting tritium-labeled dNTP, and serial dilutions of the α-CNP.

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: Paragraph title: Protein expression and purification ... The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902.

    Modification:

    Article Title: Nuclear Accumulation of S-Adenosylhomocysteine Hydrolase in Transcriptionally Active Cells during Development of Xenopus laevis
    Article Snippet: Total DNA was isolated from embryos of different stages, and the degree of methylation in CpG-containing sequences was determined by a modified nearest-neighbor analysis ( ). .. Five to 10 μg of DNA were nicked with 10 U of DNase I (Boehringer Mannheim, Mannheim, Germany) for 15 min at 37°C and 3′-end labeled with 25 μCi of [α32 P]dGTP (3000 Ci/mmol; Amersham, Braunschweig, Germany) and 10 U DNA polymerase I (Sigma, Deisenhofen, Germany) for 30 min at 15°C in a 25-μl reaction mixture containing 50 mM Tris-HCl, pH 7.5, 5.0 mM CaCl2 , and 0.04% β-mercaptoethanol.

    Crystallization Assay:

    Article Title: Crystal structure of a replicative DNA polymerase bound to the oxidized guanine lesion guanidinohydantoin
    Article Snippet: Paragraph title: Crystallization ... The exonuclease deficient DNA polymerase (RB69 exo- ) was mixed in an equimolar ratio (0.1 mM) with annealed primer template DNA and 10 mM dideoxyATP (Sigma-Aldrich).

    In Vitro:

    Article Title: Contrasting Effects of W781V and W780V Mutations in Helix N of Herpes Simplex Virus 1 and Human Cytomegalovirus DNA Polymerases on Antiviral Drug Susceptibility
    Article Snippet: The HSV-1 and HCMV DNA polymerase genes were cloned into pCITE4a (EMD BioScience, San Diego, CA) using the NdeI/NcoI and NdeI/SacI restriction sites to generate pCITE-UL30 and pCITE-UL54, respectively. .. Enzymes were expressed by an in vitro transcription-translation system using rabbit reticulocyte lysates (Promega Biosystems, Sunnyvale, CA), as previously reported ( ).

    Nick Translation:

    Article Title: Leptin protects hippocampal CA1 neurons against ischemic injury
    Article Snippet: Paragraph title: Detection of DNA fragmentation by nick-translation ... The sections were then incubated in a moist-air chamber at 37°C for 90 min with PANT reaction mixture containing 5 mM MgCl2 , 10 mM 2-mercaptoethanol, 20 μg/mL bovine serum albumin, dGTP, dCTP, and dTTP at 30 μM each, 29 μM biotinylated dATP, 1 μM dATP, and 40 U/mL Escherichia coli DNA polymerase I (Sigma) in PBS (pH 7.4).

    Sequencing:

    Article Title: Contrasting Effects of W781V and W780V Mutations in Helix N of Herpes Simplex Virus 1 and Human Cytomegalovirus DNA Polymerases on Antiviral Drug Susceptibility
    Article Snippet: The HSV-1 and HCMV DNA polymerase genes were cloned into pCITE4a (EMD BioScience, San Diego, CA) using the NdeI/NcoI and NdeI/SacI restriction sites to generate pCITE-UL30 and pCITE-UL54, respectively. .. The presence of the desired mutations was confirmed by sequencing the entire UL30 or UL54 gene in the mutated plasmids.

    Radioactivity:

    Article Title: Guanine α-carboxy nucleoside phosphonate (G-α-CNP) shows a different inhibitory kinetic profile against the DNA polymerases of human immunodeficiency virus (HIV) and herpes viruses
    Article Snippet: To perform the HCMV DNA polymerase assay, 4 μl of the TnT® reaction product was added to a 46 μl mixture to obtain 25 mM Tris.HCl pH 8.0, 100 mM (NH4 )2 SO4 (Sigma), 0.5 mM dithiothreitol, 10 mM MgCl2 , 0.2 mg/ml bovine serum albumin, 5 % glycerol (Acros, Geel, Belgium), 150 ng per μl activated calf thymus DNA (from Amersham Biosciences, Piscataway, N.J.), 100 μM of each of the three unlabeled dNTPs (GE Healthcare), and 0.5 μM of the rate-limiting tritium-labeled dNTP, and serial dilutions of the α-CNP. .. Radioactivity was determined in a Packard (Perkin Elmer, Zaventem, Belgium) Tri-Carb 2300 TR liquid scintillation counter.

    Methylation:

    Article Title: Nuclear Accumulation of S-Adenosylhomocysteine Hydrolase in Transcriptionally Active Cells during Development of Xenopus laevis
    Article Snippet: Total DNA was isolated from embryos of different stages, and the degree of methylation in CpG-containing sequences was determined by a modified nearest-neighbor analysis ( ). .. Five to 10 μg of DNA were nicked with 10 U of DNase I (Boehringer Mannheim, Mannheim, Germany) for 15 min at 37°C and 3′-end labeled with 25 μCi of [α32 P]dGTP (3000 Ci/mmol; Amersham, Braunschweig, Germany) and 10 U DNA polymerase I (Sigma, Deisenhofen, Germany) for 30 min at 15°C in a 25-μl reaction mixture containing 50 mM Tris-HCl, pH 7.5, 5.0 mM CaCl2 , and 0.04% β-mercaptoethanol.

    Mutagenesis:

    Article Title: Contrasting Effects of W781V and W780V Mutations in Helix N of Herpes Simplex Virus 1 and Human Cytomegalovirus DNA Polymerases on Antiviral Drug Susceptibility
    Article Snippet: The HSV-1 and HCMV DNA polymerase genes were cloned into pCITE4a (EMD BioScience, San Diego, CA) using the NdeI/NcoI and NdeI/SacI restriction sites to generate pCITE-UL30 and pCITE-UL54, respectively. .. The W781V and W780V mutations were introduced into the UL30 (pCITE-UL30) and UL54 (pCITE-UL54) genes, respectively, using a QuikChange site-directed mutagenesis kit (Stratagene) to produce plasmids pCITE-W781V and pCITE-W780V, respectively.

    Isolation:

    Article Title: Nuclear Accumulation of S-Adenosylhomocysteine Hydrolase in Transcriptionally Active Cells during Development of Xenopus laevis
    Article Snippet: Total DNA was isolated from embryos of different stages, and the degree of methylation in CpG-containing sequences was determined by a modified nearest-neighbor analysis ( ). .. Five to 10 μg of DNA were nicked with 10 U of DNase I (Boehringer Mannheim, Mannheim, Germany) for 15 min at 37°C and 3′-end labeled with 25 μCi of [α32 P]dGTP (3000 Ci/mmol; Amersham, Braunschweig, Germany) and 10 U DNA polymerase I (Sigma, Deisenhofen, Germany) for 30 min at 15°C in a 25-μl reaction mixture containing 50 mM Tris-HCl, pH 7.5, 5.0 mM CaCl2 , and 0.04% β-mercaptoethanol.

    Article Title: SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells
    Article Snippet: Chromatin Immunoprecipitation Assay Chromatin immunoprecipitation (ChIP) assay was performed using the EZ Magna ChIP G Chromatin Immunoprecipitation kit (Millipore, Billerica, MA, USA) using chromatin isolated from 1 × 106 cells per condition, according to the manufacturer’s protocol. .. Antibodies included anti-DNMT1 (1:500, Cell Signaling Technology), anti-IgG (Millipore), and anti-DNA polymerase II (Millipore); anti-IgG was the negative control and anti-DNA polymerase II was the positive control.

    Labeling:

    Article Title: Nuclear Accumulation of S-Adenosylhomocysteine Hydrolase in Transcriptionally Active Cells during Development of Xenopus laevis
    Article Snippet: .. Five to 10 μg of DNA were nicked with 10 U of DNase I (Boehringer Mannheim, Mannheim, Germany) for 15 min at 37°C and 3′-end labeled with 25 μCi of [α32 P]dGTP (3000 Ci/mmol; Amersham, Braunschweig, Germany) and 10 U DNA polymerase I (Sigma, Deisenhofen, Germany) for 30 min at 15°C in a 25-μl reaction mixture containing 50 mM Tris-HCl, pH 7.5, 5.0 mM CaCl2 , and 0.04% β-mercaptoethanol. .. After termination by 10 mM EDTA, nonincorporated dGTP was removed using a Sephadex G-50 minicolumn (Boehringer Mannheim).

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin. ..

    Purification:

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: Paragraph title: Protein expression and purification ... The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902.

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: .. DNA polymerase activity of extracts or purified enzyme was assayed by incubating an appropriate range of enzyme dilutions with 250 µg/ml activated (partially DNase digested) calf thymus DNA (Sigma) in KLA PCR buffer, in the presence of 50 µM each dNTP and 1 µCi [α-32 P]dCTP, at 37 or 68°C for 10 min. Aliquots of 5 µl of the samples were dotted on 3MM filter paper and washed for 3 × 15 min with 5% TCA/1% sodium pyrophosphate. .. After drying, filters were exposed to a phosphorimager screen and the intensities of the dots were quantitated with ImageQuant software.

    Polymerase Chain Reaction:

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: .. The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902. .. The gene (TK1903) encoding the large subunit of PolD (DP2) without the intein was cloned by GeneArt into pET-21a (Novagen) to generate pET-TK1903.

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: .. DNA polymerase activity of extracts or purified enzyme was assayed by incubating an appropriate range of enzyme dilutions with 250 µg/ml activated (partially DNase digested) calf thymus DNA (Sigma) in KLA PCR buffer, in the presence of 50 µM each dNTP and 1 µCi [α-32 P]dCTP, at 37 or 68°C for 10 min. Aliquots of 5 µl of the samples were dotted on 3MM filter paper and washed for 3 × 15 min with 5% TCA/1% sodium pyrophosphate. .. After drying, filters were exposed to a phosphorimager screen and the intensities of the dots were quantitated with ImageQuant software.

    Affinity Column:

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902. .. The PolD sub-units encoded by these plasmids have C-terminal His6 -tags and so were purified, after expression in Escherichia coli BL21 DE3 Rosetta cells, by Ni2+ -affinity chromatography column as previously described for the purification of PolB ( ).

    Staining:

    Article Title: A Mitosis Block Links Active Cell Cycle with Human Epidermal Differentiation and Results in Endoreplication
    Article Snippet: To test the specificity of the reaction, the DNA polymerase alpha and delta inhibitor Aphidicolin (Sigma-Aldrich, Inc.) was added in some cases to inhibit DNA replication (3 µg/ml). .. They were then washed in PBS and fixed/permeabilised in methanol (−20°C) for 10 min., washed and stained with fluorescein-labelled anti-digoxigenin (Roche) for 1 h at room temperature, washed again and mounted in Mowiol medium.

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: After electrophoresis (25 min, 25 V), air-dried and neutralized slides were stained with 2 μg/ml propidium iodide. .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin.

    Article Title: Respiratory hyperoxia reverses immunosuppression by regulating myeloid-derived suppressor cells and PD-L1 expression in a triple-negative breast cancer mouse model
    Article Snippet: Lung samples were finely minced and digested in 5 ml of an enzyme cocktail containing 1 × RPMI-1640, 2% (v/v) FBS, 0.002% (w/v) collagenase D (Sigma-Aldrich; Saint Louis, MO, USA) and 0.002% (w/v) DNA polymerase I (Sigma-Aldrich) for 1 hour at 37°C. .. After 10 to 14 days, the cells were fixed in methanol and stained with 0.03% methylene blue (Sigma-Aldrich).

    Mouse Assay:

    Article Title: Respiratory hyperoxia reverses immunosuppression by regulating myeloid-derived suppressor cells and PD-L1 expression in a triple-negative breast cancer mouse model
    Article Snippet: In brief, on day 7, 14, or 21, 4T1 tumor-bearing mice were sacrificed, and their lungs were harvested. .. Lung samples were finely minced and digested in 5 ml of an enzyme cocktail containing 1 × RPMI-1640, 2% (v/v) FBS, 0.002% (w/v) collagenase D (Sigma-Aldrich; Saint Louis, MO, USA) and 0.002% (w/v) DNA polymerase I (Sigma-Aldrich) for 1 hour at 37°C.

    Chromatin Immunoprecipitation:

    Article Title: SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells
    Article Snippet: Paragraph title: 4.9. Chromatin Immunoprecipitation Assay ... Antibodies included anti-DNMT1 (1:500, Cell Signaling Technology), anti-IgG (Millipore), and anti-DNA polymerase II (Millipore); anti-IgG was the negative control and anti-DNA polymerase II was the positive control.

    Plasmid Preparation:

    Article Title: Guanine α-carboxy nucleoside phosphonate (G-α-CNP) shows a different inhibitory kinetic profile against the DNA polymerases of human immunodeficiency virus (HIV) and herpes viruses
    Article Snippet: The plasmid was added (at 10 ng per μl volume) to the TnT® mix containing 0.5 mM MgCl2 and 10 mM potassium acetate (Janssen Chimica, Beerse, Belgium), and the mixture was incubated for 3 hours at 30°C. .. To perform the HCMV DNA polymerase assay, 4 μl of the TnT® reaction product was added to a 46 μl mixture to obtain 25 mM Tris.HCl pH 8.0, 100 mM (NH4 )2 SO4 (Sigma), 0.5 mM dithiothreitol, 10 mM MgCl2 , 0.2 mg/ml bovine serum albumin, 5 % glycerol (Acros, Geel, Belgium), 150 ng per μl activated calf thymus DNA (from Amersham Biosciences, Piscataway, N.J.), 100 μM of each of the three unlabeled dNTPs (GE Healthcare), and 0.5 μM of the rate-limiting tritium-labeled dNTP, and serial dilutions of the α-CNP.

    Software:

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: Average comet tail moment was scored for duplicate slides (50 cells/slide) in randomly selected fields, and automated calculations were performed using Comet Assay II software (Perceptive Instruments, Suffolk, UK). .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin.

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: DNA polymerase activity of extracts or purified enzyme was assayed by incubating an appropriate range of enzyme dilutions with 250 µg/ml activated (partially DNase digested) calf thymus DNA (Sigma) in KLA PCR buffer, in the presence of 50 µM each dNTP and 1 µCi [α-32 P]dCTP, at 37 or 68°C for 10 min. Aliquots of 5 µl of the samples were dotted on 3MM filter paper and washed for 3 × 15 min with 5% TCA/1% sodium pyrophosphate. .. After drying, filters were exposed to a phosphorimager screen and the intensities of the dots were quantitated with ImageQuant software.

    Negative Control:

    Article Title: SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells
    Article Snippet: .. Antibodies included anti-DNMT1 (1:500, Cell Signaling Technology), anti-IgG (Millipore), and anti-DNA polymerase II (Millipore); anti-IgG was the negative control and anti-DNA polymerase II was the positive control. .. Transfection With Short Hairpin RNA Cells were transfected with short hairpin RNA (shRNA) by using Lipofectamine 2000 (Invitrogen), following the procedure recommended by the manufacturer. shRNA targeting of CLDN6 (5’-GTGCAAGGTGTACGACTCA-3’) and a negative control shRNA were purchased from GeneChem Co. Ltd.

    Positron Emission Tomography:

    Article Title: Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability
    Article Snippet: .. The gene (TK1902) encoding the small subunit of DNA polymerase D (DP1) was PCR amplified from genomic DNA and cloned into pET-21a (Novagen) to generate pET-TK1902. .. The gene (TK1903) encoding the large subunit of PolD (DP2) without the intein was cloned by GeneArt into pET-21a (Novagen) to generate pET-TK1903.

    In Situ:

    Article Title: A Mitosis Block Links Active Cell Cycle with Human Epidermal Differentiation and Results in Endoreplication
    Article Snippet: Paragraph title: In situ DNA replication ... To test the specificity of the reaction, the DNA polymerase alpha and delta inhibitor Aphidicolin (Sigma-Aldrich, Inc.) was added in some cases to inhibit DNA replication (3 µg/ml).

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: In Situ DNA Strand Break Detection —A DNA polymerase I-mediated labeling assay was performed as described previously with minor modifications ( ). .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin.

    Incubation:

    Article Title: Leptin protects hippocampal CA1 neurons against ischemic injury
    Article Snippet: .. The sections were then incubated in a moist-air chamber at 37°C for 90 min with PANT reaction mixture containing 5 mM MgCl2 , 10 mM 2-mercaptoethanol, 20 μg/mL bovine serum albumin, dGTP, dCTP, and dTTP at 30 μM each, 29 μM biotinylated dATP, 1 μM dATP, and 40 U/mL Escherichia coli DNA polymerase I (Sigma) in PBS (pH 7.4). ..

    Article Title: Guanine α-carboxy nucleoside phosphonate (G-α-CNP) shows a different inhibitory kinetic profile against the DNA polymerases of human immunodeficiency virus (HIV) and herpes viruses
    Article Snippet: The plasmid was added (at 10 ng per μl volume) to the TnT® mix containing 0.5 mM MgCl2 and 10 mM potassium acetate (Janssen Chimica, Beerse, Belgium), and the mixture was incubated for 3 hours at 30°C. .. To perform the HCMV DNA polymerase assay, 4 μl of the TnT® reaction product was added to a 46 μl mixture to obtain 25 mM Tris.HCl pH 8.0, 100 mM (NH4 )2 SO4 (Sigma), 0.5 mM dithiothreitol, 10 mM MgCl2 , 0.2 mg/ml bovine serum albumin, 5 % glycerol (Acros, Geel, Belgium), 150 ng per μl activated calf thymus DNA (from Amersham Biosciences, Piscataway, N.J.), 100 μM of each of the three unlabeled dNTPs (GE Healthcare), and 0.5 μM of the rate-limiting tritium-labeled dNTP, and serial dilutions of the α-CNP.

    Article Title: A Mitosis Block Links Active Cell Cycle with Human Epidermal Differentiation and Results in Endoreplication
    Article Snippet: Skin sections were then thawed and immediately were overlaid with 40 µl replication buffer, prepared with 30 µl of incubation buffer (0,25 M sucrose, 75 mM NaCl, 0,5 mM spermidin, 0,15 mM, spermin, 3% BSA) and 10 µl nucleotide mix (40 mM Khepes, 7 mM MgCl2, 0,1 mM dCTP, dATP, DGTP, rUTP, rCTP, rGTP, 60 mM ATP, 50 µM digoxigenin-UTP, 0,5 mM DTT, 80 mM creatin phosphate, 10 µg/ml phosphocreatin kinase). .. To test the specificity of the reaction, the DNA polymerase alpha and delta inhibitor Aphidicolin (Sigma-Aldrich, Inc.) was added in some cases to inhibit DNA replication (3 µg/ml).

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin. ..

    Apoptosis Assay:

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin. .. Apoptosis Assay —Equivalent numbers of indicated cells were plated and allowed to settle for 24 h at 50% confluence.

    Thin Layer Chromatography:

    Article Title: Nuclear Accumulation of S-Adenosylhomocysteine Hydrolase in Transcriptionally Active Cells during Development of Xenopus laevis
    Article Snippet: Five to 10 μg of DNA were nicked with 10 U of DNase I (Boehringer Mannheim, Mannheim, Germany) for 15 min at 37°C and 3′-end labeled with 25 μCi of [α32 P]dGTP (3000 Ci/mmol; Amersham, Braunschweig, Germany) and 10 U DNA polymerase I (Sigma, Deisenhofen, Germany) for 30 min at 15°C in a 25-μl reaction mixture containing 50 mM Tris-HCl, pH 7.5, 5.0 mM CaCl2 , and 0.04% β-mercaptoethanol. .. The resulting desoxynucleoside 3′-monophosphates were applied to cellulose TLC sheets (Machery-Nagel, Düren, Germany) and separated in two dimensions ( ).

    Lysis:

    Article Title: E2F1 Regulates the Base Excision Repair Gene XRCC1 and Promotes DNA Repair *
    Article Snippet: Slides were submerged in lysis buffer (2.5 m NaCl, 0.1 m EDTA, 10 m m Tris-Cl (pH 7.0), 1% Triton X-100, 1% DMSO) for 1 h, washed with nanopure H2 O, and incubated for 45 min in alkaline electrophoresis buffer (50 m m NaOH, 1 m m EDTA, 1% DMSO, pH 12.8). .. Cells were rinsed in phosphate-buffered saline, permeabilized with 1% Triton X-100 for 20 min, and then incubated in a moist air chamber at 37 °C for 90 min in a labeling mixture containing 10 μ m each dGTP, dATP, and dCTP and 7 μ m dTTP, 3 μ m FITC-dUTP, 20 units/ml Escherichia coli DNA polymerase I (Sigma) in reaction buffer containing 5 m m MgCl2 , 10 m m 2-mercaptoethanol, and 20 μg/ml bovine serum albumin.

    Activity Assay:

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: .. DNA polymerase activity of extracts or purified enzyme was assayed by incubating an appropriate range of enzyme dilutions with 250 µg/ml activated (partially DNase digested) calf thymus DNA (Sigma) in KLA PCR buffer, in the presence of 50 µM each dNTP and 1 µCi [α-32 P]dCTP, at 37 or 68°C for 10 min. Aliquots of 5 µl of the samples were dotted on 3MM filter paper and washed for 3 × 15 min with 5% TCA/1% sodium pyrophosphate. .. After drying, filters were exposed to a phosphorimager screen and the intensities of the dots were quantitated with ImageQuant software.

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  • 99
    Millipore anti dna polymerase ii
    <t>DNA</t> methyltransferase 1 <t>(DNMT1)-mediated</t> upregulation of CLDN6 expression by SB431542. ( A , B ) Real-time polymerase chain reaction (RT-PCR) and immunoblot analysis of DNMT1 and CLDN6, and densitometric analysis of relative gene expression levels after normalization to loading controls GAPDH and β-actin are presented. ( C ) DNMT1 activity assays. ( D ) Methylation-specific PCR (MSP) analysis of CpG island of CLDN6 promoter using bisulfite-treated genomic DNA isolated from MCF-7 and SKBR-3 cells. ( E ) CpG island methylation within the CLDN6 promoter region was measured by bisulfite sequencing in SKBR-3 cells. “Me” stands for methylated, and “U” stands for unmethylated. ( F ) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay to detect the binding of DNMT1 to the promoter of CLDN6 (** p
    Anti Dna Polymerase Ii, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti dna polymerase ii/product/Millipore
    Average 99 stars, based on 1 article reviews
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    anti dna polymerase ii - by Bioz Stars, 2020-04
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    94
    Millipore mouse anti rna pol ii
    THE1B 5′ insertion site creates novel binding site for transcription factor DLX3. (A) Top left, sequence of 5′ insertion site of THE1B near CRH , predicted DLX3 binding site shown in red. Lower left, electrophoretic mobility shift assay demonstrating binding of DLX3-DDK to positive control (JRE probe) and the 5′ insertion site of THE1B (THE1B 5′ probe). The red arrow denotes the band formed by DLX3-DDK binding to labeled DNA probe; this binding is disrupted with the addition of anti-DDK but not isotype control mouse <t>IgG.</t> Top right, same sequence as left, predicted GATA2 binding site shown in red. Lower right, GATA2-DDK fails to bind to the 5′ insertion site of THE1B (THE1B 5′ probe) while binding positive control (TCR probe). Blue arrow denotes the supershifted band of anti-DDK, GATA2-DDK, and TCR probe. (B) DNA binding motif of transcription factor DLX3 [ 46 ]. This DLX3 binding site is formed by the insertion of THE1B (right of black bar) into the genome (left of black bar). (C) ChIP for DLX3 (black bars) and <t>RNA</t> polymerase II (white bar) with quantitative real-time PCR. DLX3 is significantly associated with the 5′ end of THE1B and positive control (JRE) in human term placenta. ChIP-qPCR data were normalized to the IgG control for each target and presented relative to negative control (JRE distal). ( n = 3 for all, P
    Mouse Anti Rna Pol Ii, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti rna pol ii/product/Millipore
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    Price from $9.99 to $1999.99
    mouse anti rna pol ii - by Bioz Stars, 2020-04
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    86
    Millipore monoclonal mouse anti human rna polymerase ii
    Omeprazole (Ome) decreases IL-4-stimulated <t>RNA</t> Pol II binding to the endogenous eotaxin-3 promoter in (A) EoE1-T and (B) EoE2-T cells. Data are the means ± SEM of 3 separate experiments. **, p≤0.01 compared to IL-4 stimulation; ***, p≤0.001 compared to IL-4 stimulation Isotype matched IgG served as a control. Representative experiment demonstrating that omeprazole reduces the levels of IL-4 stimulated <t>H3K4me3</t> bound to the endogenous eotaxin-3 promoter in (C) EoE1-T and (D) EoE2-T cells. Isotype matched IgG served as a control. Depicted is one of 3 separate experiments. M, marker.
    Monoclonal Mouse Anti Human Rna Polymerase Ii, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    DNA methyltransferase 1 (DNMT1)-mediated upregulation of CLDN6 expression by SB431542. ( A , B ) Real-time polymerase chain reaction (RT-PCR) and immunoblot analysis of DNMT1 and CLDN6, and densitometric analysis of relative gene expression levels after normalization to loading controls GAPDH and β-actin are presented. ( C ) DNMT1 activity assays. ( D ) Methylation-specific PCR (MSP) analysis of CpG island of CLDN6 promoter using bisulfite-treated genomic DNA isolated from MCF-7 and SKBR-3 cells. ( E ) CpG island methylation within the CLDN6 promoter region was measured by bisulfite sequencing in SKBR-3 cells. “Me” stands for methylated, and “U” stands for unmethylated. ( F ) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay to detect the binding of DNMT1 to the promoter of CLDN6 (** p

    Journal: International Journal of Molecular Sciences

    Article Title: SMAD2 Inactivation Inhibits CLDN6 Methylation to Suppress Migration and Invasion of Breast Cancer Cells

    doi: 10.3390/ijms18091863

    Figure Lengend Snippet: DNA methyltransferase 1 (DNMT1)-mediated upregulation of CLDN6 expression by SB431542. ( A , B ) Real-time polymerase chain reaction (RT-PCR) and immunoblot analysis of DNMT1 and CLDN6, and densitometric analysis of relative gene expression levels after normalization to loading controls GAPDH and β-actin are presented. ( C ) DNMT1 activity assays. ( D ) Methylation-specific PCR (MSP) analysis of CpG island of CLDN6 promoter using bisulfite-treated genomic DNA isolated from MCF-7 and SKBR-3 cells. ( E ) CpG island methylation within the CLDN6 promoter region was measured by bisulfite sequencing in SKBR-3 cells. “Me” stands for methylated, and “U” stands for unmethylated. ( F ) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay to detect the binding of DNMT1 to the promoter of CLDN6 (** p

    Article Snippet: Antibodies included anti-DNMT1 (1:500, Cell Signaling Technology), anti-IgG (Millipore), and anti-DNA polymerase II (Millipore); anti-IgG was the negative control and anti-DNA polymerase II was the positive control.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Methylation, Polymerase Chain Reaction, Isolation, Methylation Sequencing, Chromatin Immunoprecipitation, Binding Assay

    THE1B 5′ insertion site creates novel binding site for transcription factor DLX3. (A) Top left, sequence of 5′ insertion site of THE1B near CRH , predicted DLX3 binding site shown in red. Lower left, electrophoretic mobility shift assay demonstrating binding of DLX3-DDK to positive control (JRE probe) and the 5′ insertion site of THE1B (THE1B 5′ probe). The red arrow denotes the band formed by DLX3-DDK binding to labeled DNA probe; this binding is disrupted with the addition of anti-DDK but not isotype control mouse IgG. Top right, same sequence as left, predicted GATA2 binding site shown in red. Lower right, GATA2-DDK fails to bind to the 5′ insertion site of THE1B (THE1B 5′ probe) while binding positive control (TCR probe). Blue arrow denotes the supershifted band of anti-DDK, GATA2-DDK, and TCR probe. (B) DNA binding motif of transcription factor DLX3 [ 46 ]. This DLX3 binding site is formed by the insertion of THE1B (right of black bar) into the genome (left of black bar). (C) ChIP for DLX3 (black bars) and RNA polymerase II (white bar) with quantitative real-time PCR. DLX3 is significantly associated with the 5′ end of THE1B and positive control (JRE) in human term placenta. ChIP-qPCR data were normalized to the IgG control for each target and presented relative to negative control (JRE distal). ( n = 3 for all, P

    Journal: PLoS Biology

    Article Title: Anthropoid primate–specific retroviral element THE1B controls expression of CRH in placenta and alters gestation length

    doi: 10.1371/journal.pbio.2006337

    Figure Lengend Snippet: THE1B 5′ insertion site creates novel binding site for transcription factor DLX3. (A) Top left, sequence of 5′ insertion site of THE1B near CRH , predicted DLX3 binding site shown in red. Lower left, electrophoretic mobility shift assay demonstrating binding of DLX3-DDK to positive control (JRE probe) and the 5′ insertion site of THE1B (THE1B 5′ probe). The red arrow denotes the band formed by DLX3-DDK binding to labeled DNA probe; this binding is disrupted with the addition of anti-DDK but not isotype control mouse IgG. Top right, same sequence as left, predicted GATA2 binding site shown in red. Lower right, GATA2-DDK fails to bind to the 5′ insertion site of THE1B (THE1B 5′ probe) while binding positive control (TCR probe). Blue arrow denotes the supershifted band of anti-DDK, GATA2-DDK, and TCR probe. (B) DNA binding motif of transcription factor DLX3 [ 46 ]. This DLX3 binding site is formed by the insertion of THE1B (right of black bar) into the genome (left of black bar). (C) ChIP for DLX3 (black bars) and RNA polymerase II (white bar) with quantitative real-time PCR. DLX3 is significantly associated with the 5′ end of THE1B and positive control (JRE) in human term placenta. ChIP-qPCR data were normalized to the IgG control for each target and presented relative to negative control (JRE distal). ( n = 3 for all, P

    Article Snippet: Mouse anti-RNA pol II (Millipore 05-623B) and Control Mouse IgG antibodies (Millipore 12-371B) were used as a positive and negative control according to EZ-ChIP protocol.

    Techniques: Binding Assay, Sequencing, Electrophoretic Mobility Shift Assay, Positive Control, Labeling, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control

    Omeprazole (Ome) decreases IL-4-stimulated RNA Pol II binding to the endogenous eotaxin-3 promoter in (A) EoE1-T and (B) EoE2-T cells. Data are the means ± SEM of 3 separate experiments. **, p≤0.01 compared to IL-4 stimulation; ***, p≤0.001 compared to IL-4 stimulation Isotype matched IgG served as a control. Representative experiment demonstrating that omeprazole reduces the levels of IL-4 stimulated H3K4me3 bound to the endogenous eotaxin-3 promoter in (C) EoE1-T and (D) EoE2-T cells. Isotype matched IgG served as a control. Depicted is one of 3 separate experiments. M, marker.

    Journal: PLoS ONE

    Article Title: Omeprazole Blocks STAT6 Binding to the Eotaxin-3 Promoter in Eosinophilic Esophagitis Cells

    doi: 10.1371/journal.pone.0050037

    Figure Lengend Snippet: Omeprazole (Ome) decreases IL-4-stimulated RNA Pol II binding to the endogenous eotaxin-3 promoter in (A) EoE1-T and (B) EoE2-T cells. Data are the means ± SEM of 3 separate experiments. **, p≤0.01 compared to IL-4 stimulation; ***, p≤0.001 compared to IL-4 stimulation Isotype matched IgG served as a control. Representative experiment demonstrating that omeprazole reduces the levels of IL-4 stimulated H3K4me3 bound to the endogenous eotaxin-3 promoter in (C) EoE1-T and (D) EoE2-T cells. Isotype matched IgG served as a control. Depicted is one of 3 separate experiments. M, marker.

    Article Snippet: The sheared chromatin was cleared by centrifugation and the supernatant (500 µg) was immunoprecipitated overnight at 4°C with 1.6 µg of polyclonal rabbit anti-human STAT6 (Santa Cruz) or anti-human Histone H3 trimethyl K4 (H3K4me3, Abcam) or monoclonal mouse anti-human RNA Polymerase II (Millipore); rabbit IgG or mouse IgG were used as isotype controls.

    Techniques: Binding Assay, Marker