Structured Review

Promega exonuclease iii digestion
Specific activation of MAPK by ΔB-Raf:ER increases expression of <t>PEG-3.</t> B1TD cells were infected with either a control recombinant adenovirus (CMV) or a recombinant adenovirus to express ΔB-Raf:ER. Twenty-four hours after infection, cells were treated with 200 nM 4-hydroxytamoxifen (TAM) and either vehicle control (VEH, DMSO) or 50 µM PD98059. Six hours after treatment, cells were washed with PBS and frozen. ( A ) Cells were lysed and prepared for immune complex kinase assays to determine MAPK activity, as described in Materials and Methods. Data are the means of <t>three</t> separate experiments ± SEM. ( B ) Cells were lysed in preparation for SDS–PAGE, followed by immunoblotting to determine PEG-3 protein expression levels. A representative experiment is shown n = 3. Lane 1, Ad. vec ; lane 2, Ad. vec + TAM; lane 3, Ad. vec + TAM + PD98059; lane 4, Ad.ΔB-Raf:ER; lane 5, Ad.ΔB-Raf:ER + TAM; lane 6, Ad.ΔB-Raf:ER + TAM + PD98059.
Exonuclease Iii Digestion, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/exonuclease iii digestion/product/Promega
Average 88 stars, based on 8 article reviews
Price from $9.99 to $1999.99
exonuclease iii digestion - by Bioz Stars, 2020-09
88/100 stars

Images

1) Product Images from "PEA3 sites within the progression elevated gene-3 (PEG-3) promoter and mitogen-activated protein kinase contribute to differential PEG-3 expression in Ha-ras and v-raf oncogene transformed rat embryo cells"

Article Title: PEA3 sites within the progression elevated gene-3 (PEG-3) promoter and mitogen-activated protein kinase contribute to differential PEG-3 expression in Ha-ras and v-raf oncogene transformed rat embryo cells

Journal: Nucleic Acids Research

doi:

Specific activation of MAPK by ΔB-Raf:ER increases expression of PEG-3. B1TD cells were infected with either a control recombinant adenovirus (CMV) or a recombinant adenovirus to express ΔB-Raf:ER. Twenty-four hours after infection, cells were treated with 200 nM 4-hydroxytamoxifen (TAM) and either vehicle control (VEH, DMSO) or 50 µM PD98059. Six hours after treatment, cells were washed with PBS and frozen. ( A ) Cells were lysed and prepared for immune complex kinase assays to determine MAPK activity, as described in Materials and Methods. Data are the means of three separate experiments ± SEM. ( B ) Cells were lysed in preparation for SDS–PAGE, followed by immunoblotting to determine PEG-3 protein expression levels. A representative experiment is shown n = 3. Lane 1, Ad. vec ; lane 2, Ad. vec + TAM; lane 3, Ad. vec + TAM + PD98059; lane 4, Ad.ΔB-Raf:ER; lane 5, Ad.ΔB-Raf:ER + TAM; lane 6, Ad.ΔB-Raf:ER + TAM + PD98059.
Figure Legend Snippet: Specific activation of MAPK by ΔB-Raf:ER increases expression of PEG-3. B1TD cells were infected with either a control recombinant adenovirus (CMV) or a recombinant adenovirus to express ΔB-Raf:ER. Twenty-four hours after infection, cells were treated with 200 nM 4-hydroxytamoxifen (TAM) and either vehicle control (VEH, DMSO) or 50 µM PD98059. Six hours after treatment, cells were washed with PBS and frozen. ( A ) Cells were lysed and prepared for immune complex kinase assays to determine MAPK activity, as described in Materials and Methods. Data are the means of three separate experiments ± SEM. ( B ) Cells were lysed in preparation for SDS–PAGE, followed by immunoblotting to determine PEG-3 protein expression levels. A representative experiment is shown n = 3. Lane 1, Ad. vec ; lane 2, Ad. vec + TAM; lane 3, Ad. vec + TAM + PD98059; lane 4, Ad.ΔB-Raf:ER; lane 5, Ad.ΔB-Raf:ER + TAM; lane 6, Ad.ΔB-Raf:ER + TAM + PD98059.

Techniques Used: Activation Assay, Expressing, Infection, Recombinant, Immune Complex Kinase Assay, Activity Assay, SDS Page

2) Product Images from "O6-Methylguanine induces altered proteins at the level of transcription in human cells"

Article Title: O6-Methylguanine induces altered proteins at the level of transcription in human cells

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkq706

qRT 2 -PCR indicates presence of RFPm mRNA in the absence of RFP fluorescence. Although there is no red fluorescence in cells transfected with pRFPm, RNA quantification shows there is no significant difference between RFP mRNA levels in cells transfected with pRFPwt, pRFPm, pRFPm- O 6 meG and pRFPm- O 6 meG plus 10 µM O 6 -benzylguanine. Additionally, treatment with 50 µM O 6 -benzylguanine significantly affected mRNA levels in cells transfected with pRFPm, and pRFPm- O 6 meG. Significance was determined by a two-sample Student’s t -test which compared the RFP–GFP mRNA abundance ratio from untreated pRwt transfected cells to the ratio observed under each of the experimental conditions in at least three independent experiments (* P
Figure Legend Snippet: qRT 2 -PCR indicates presence of RFPm mRNA in the absence of RFP fluorescence. Although there is no red fluorescence in cells transfected with pRFPm, RNA quantification shows there is no significant difference between RFP mRNA levels in cells transfected with pRFPwt, pRFPm, pRFPm- O 6 meG and pRFPm- O 6 meG plus 10 µM O 6 -benzylguanine. Additionally, treatment with 50 µM O 6 -benzylguanine significantly affected mRNA levels in cells transfected with pRFPm, and pRFPm- O 6 meG. Significance was determined by a two-sample Student’s t -test which compared the RFP–GFP mRNA abundance ratio from untreated pRwt transfected cells to the ratio observed under each of the experimental conditions in at least three independent experiments (* P

Techniques Used: Polymerase Chain Reaction, Fluorescence, Transfection

3) Product Images from "IFN-? Attenuates Hypoxia-Inducible Factor (HIF) Activity in Intestinal Epithelial Cells through Transcriptional Repression of HIF-1?"

Article Title: IFN-? Attenuates Hypoxia-Inducible Factor (HIF) Activity in Intestinal Epithelial Cells through Transcriptional Repression of HIF-1?

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi: 10.4049/jimmunol.1001442

Repression of HIF-1β promoter activity by IFN-γ. A , Cloned HIF-1β promoter and series of truncated promoter-luciferase reporter constructs generated using exonuclease III digestion. Relative positions of each clone and major TSSs
Figure Legend Snippet: Repression of HIF-1β promoter activity by IFN-γ. A , Cloned HIF-1β promoter and series of truncated promoter-luciferase reporter constructs generated using exonuclease III digestion. Relative positions of each clone and major TSSs

Techniques Used: Activity Assay, Clone Assay, Luciferase, Construct, Generated

4) Product Images from "Insights into glutamate transport regulation in human astrocytes: Cloning of the promoter for excitatory amino acid transporter 2 (EAAT2)"

Article Title: Insights into glutamate transport regulation in human astrocytes: Cloning of the promoter for excitatory amino acid transporter 2 (EAAT2)

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0136555100

Effect of pharmacological inhibitors on EAAT2-Prom activity, and mRNA and protein expression in PHFA after various treatment protocols. ( A–C ) EAAT2-Prom activity in PHFA either untreated (−) or treated with EGF (30 ng/ml) ( A ), bromo-cAMP (250 μM) ( B ), or TNF-α (200 units/ml) ( C ) in the absence (−) or presence (+) of KT5720 (5 μM), AG1478 (1 μM), PDTC (100 μM), wortmanin (WRT) (100 nM), or PD98049 (PD) (50 μM). The duration of the assay was 4 days. ( D ) Effect of the various treatment protocols on EAAT2 and GAPDH mRNA levels. ( Upper ) Relative EAAT2 RNA expression versus GAPDH expression compared with control untreated or treated samples based on scanning of autoradiograms. ( Lower ) Actual Northern blots. ( E ) Effect of the various treatment protocols on EAAT2 and ACTIN protein levels. ( Upper ) Relative EAAT2 protein expression versus ACTIN expression compared with control untreated or treated samples based on scanning of autoradiograms. Individual experiments were performed three times with triplicate samples, and SD from the mean is presented.
Figure Legend Snippet: Effect of pharmacological inhibitors on EAAT2-Prom activity, and mRNA and protein expression in PHFA after various treatment protocols. ( A–C ) EAAT2-Prom activity in PHFA either untreated (−) or treated with EGF (30 ng/ml) ( A ), bromo-cAMP (250 μM) ( B ), or TNF-α (200 units/ml) ( C ) in the absence (−) or presence (+) of KT5720 (5 μM), AG1478 (1 μM), PDTC (100 μM), wortmanin (WRT) (100 nM), or PD98049 (PD) (50 μM). The duration of the assay was 4 days. ( D ) Effect of the various treatment protocols on EAAT2 and GAPDH mRNA levels. ( Upper ) Relative EAAT2 RNA expression versus GAPDH expression compared with control untreated or treated samples based on scanning of autoradiograms. ( Lower ) Actual Northern blots. ( E ) Effect of the various treatment protocols on EAAT2 and ACTIN protein levels. ( Upper ) Relative EAAT2 protein expression versus ACTIN expression compared with control untreated or treated samples based on scanning of autoradiograms. Individual experiments were performed three times with triplicate samples, and SD from the mean is presented.

Techniques Used: Activity Assay, Expressing, RNA Expression, Northern Blot

Related Articles

Sequencing:

Article Title: IFN-? Attenuates Hypoxia-Inducible Factor (HIF) Activity in Intestinal Epithelial Cells through Transcriptional Repression of HIF-1?
Article Snippet: .. Sequential truncations of the cloned HIF-1β promoter sequence were generated by exonuclease III digestion (Erase-a-Base; Promega). .. Transfection of Caco-2 and HeLa cells was carried out using FuGENE 6 (Roche Diagnostics, Indianapolis, IN), as directed by the manufacturer.

Clone Assay:

Article Title: IFN-? Attenuates Hypoxia-Inducible Factor (HIF) Activity in Intestinal Epithelial Cells through Transcriptional Repression of HIF-1?
Article Snippet: .. Sequential truncations of the cloned HIF-1β promoter sequence were generated by exonuclease III digestion (Erase-a-Base; Promega). .. Transfection of Caco-2 and HeLa cells was carried out using FuGENE 6 (Roche Diagnostics, Indianapolis, IN), as directed by the manufacturer.

Generated:

Article Title: IFN-? Attenuates Hypoxia-Inducible Factor (HIF) Activity in Intestinal Epithelial Cells through Transcriptional Repression of HIF-1?
Article Snippet: .. Sequential truncations of the cloned HIF-1β promoter sequence were generated by exonuclease III digestion (Erase-a-Base; Promega). .. Transfection of Caco-2 and HeLa cells was carried out using FuGENE 6 (Roche Diagnostics, Indianapolis, IN), as directed by the manufacturer.

Construct:

Article Title: SP PROTEINS AND RUNX2 MEDIATE REGULATION OF MATRIX GLA PROTEIN (MGP) EXPRESSION BY PARATHYROID HORMONE
Article Snippet: .. 5'-deletion constructs were created by exonuclease III digestion of the pMGP-748-luc linearized with SacI and XhoI , using the Erase-a-Base kit (Promega) as recommended by the manufacturer. .. Point mutations in the pMGP-luc reporter plasmids were introduced using the QuikChange Site-Directed Mutagenesis Kit (Stratagene) according to the manufacturer's instructions.

Article Title: Interdependent Expression of the ccoNOQP-rdxBHIS Loci in Rhodobacter sphaeroides 2.4.1
Article Snippet: .. The 5′-deletion constructs −380 and −112 were created by exonuclease III digestion of pJR194 linearized with Bam HI and Sac I by using the Erase-A-Base system (Promega) according to the manufacturer's instructions. ..

Polymerase Chain Reaction:

Article Title: O6-Methylguanine induces altered proteins at the level of transcription in human cells
Article Snippet: .. An aliquot of the digested DNA containing 0.05 ng (after DpnI digestion) or 0.008 ng (after DpnI and exonuclease III digestion) was used in a PCR reaction (GoTaq green master mix, Promega) containing primers (RdF: 5′-CAGCGGCCCTTCTCTCTTA-3′; RR: 5′-CGCTACAGGAACAGGTGGTG-3′) whose product spans 7 DpnI sites. .. Fluorescence microscopy For fluorescence microscopy, 2 × 105 cells were plated on collagen type I coated four-well culture slides (BD BioCoat, BD Biosciences) and transfected 24 h later with 1 µg plasmid as described above.

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    Promega exonuclease iii digestion
    Specific activation of MAPK by ΔB-Raf:ER increases expression of <t>PEG-3.</t> B1TD cells were infected with either a control recombinant adenovirus (CMV) or a recombinant adenovirus to express ΔB-Raf:ER. Twenty-four hours after infection, cells were treated with 200 nM 4-hydroxytamoxifen (TAM) and either vehicle control (VEH, DMSO) or 50 µM PD98059. Six hours after treatment, cells were washed with PBS and frozen. ( A ) Cells were lysed and prepared for immune complex kinase assays to determine MAPK activity, as described in Materials and Methods. Data are the means of <t>three</t> separate experiments ± SEM. ( B ) Cells were lysed in preparation for SDS–PAGE, followed by immunoblotting to determine PEG-3 protein expression levels. A representative experiment is shown n = 3. Lane 1, Ad. vec ; lane 2, Ad. vec + TAM; lane 3, Ad. vec + TAM + PD98059; lane 4, Ad.ΔB-Raf:ER; lane 5, Ad.ΔB-Raf:ER + TAM; lane 6, Ad.ΔB-Raf:ER + TAM + PD98059.
    Exonuclease Iii Digestion, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/exonuclease iii digestion/product/Promega
    Average 88 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    exonuclease iii digestion - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    Image Search Results


    Specific activation of MAPK by ΔB-Raf:ER increases expression of PEG-3. B1TD cells were infected with either a control recombinant adenovirus (CMV) or a recombinant adenovirus to express ΔB-Raf:ER. Twenty-four hours after infection, cells were treated with 200 nM 4-hydroxytamoxifen (TAM) and either vehicle control (VEH, DMSO) or 50 µM PD98059. Six hours after treatment, cells were washed with PBS and frozen. ( A ) Cells were lysed and prepared for immune complex kinase assays to determine MAPK activity, as described in Materials and Methods. Data are the means of three separate experiments ± SEM. ( B ) Cells were lysed in preparation for SDS–PAGE, followed by immunoblotting to determine PEG-3 protein expression levels. A representative experiment is shown n = 3. Lane 1, Ad. vec ; lane 2, Ad. vec + TAM; lane 3, Ad. vec + TAM + PD98059; lane 4, Ad.ΔB-Raf:ER; lane 5, Ad.ΔB-Raf:ER + TAM; lane 6, Ad.ΔB-Raf:ER + TAM + PD98059.

    Journal: Nucleic Acids Research

    Article Title: PEA3 sites within the progression elevated gene-3 (PEG-3) promoter and mitogen-activated protein kinase contribute to differential PEG-3 expression in Ha-ras and v-raf oncogene transformed rat embryo cells

    doi:

    Figure Lengend Snippet: Specific activation of MAPK by ΔB-Raf:ER increases expression of PEG-3. B1TD cells were infected with either a control recombinant adenovirus (CMV) or a recombinant adenovirus to express ΔB-Raf:ER. Twenty-four hours after infection, cells were treated with 200 nM 4-hydroxytamoxifen (TAM) and either vehicle control (VEH, DMSO) or 50 µM PD98059. Six hours after treatment, cells were washed with PBS and frozen. ( A ) Cells were lysed and prepared for immune complex kinase assays to determine MAPK activity, as described in Materials and Methods. Data are the means of three separate experiments ± SEM. ( B ) Cells were lysed in preparation for SDS–PAGE, followed by immunoblotting to determine PEG-3 protein expression levels. A representative experiment is shown n = 3. Lane 1, Ad. vec ; lane 2, Ad. vec + TAM; lane 3, Ad. vec + TAM + PD98059; lane 4, Ad.ΔB-Raf:ER; lane 5, Ad.ΔB-Raf:ER + TAM; lane 6, Ad.ΔB-Raf:ER + TAM + PD98059.

    Article Snippet: 5′ Deletion mutations in the FL-PEG-Prom were made with exonuclease III digestion using the Erase-A-Base System (Promega).

    Techniques: Activation Assay, Expressing, Infection, Recombinant, Immune Complex Kinase Assay, Activity Assay, SDS Page

    qRT 2 -PCR indicates presence of RFPm mRNA in the absence of RFP fluorescence. Although there is no red fluorescence in cells transfected with pRFPm, RNA quantification shows there is no significant difference between RFP mRNA levels in cells transfected with pRFPwt, pRFPm, pRFPm- O 6 meG and pRFPm- O 6 meG plus 10 µM O 6 -benzylguanine. Additionally, treatment with 50 µM O 6 -benzylguanine significantly affected mRNA levels in cells transfected with pRFPm, and pRFPm- O 6 meG. Significance was determined by a two-sample Student’s t -test which compared the RFP–GFP mRNA abundance ratio from untreated pRwt transfected cells to the ratio observed under each of the experimental conditions in at least three independent experiments (* P

    Journal: Nucleic Acids Research

    Article Title: O6-Methylguanine induces altered proteins at the level of transcription in human cells

    doi: 10.1093/nar/gkq706

    Figure Lengend Snippet: qRT 2 -PCR indicates presence of RFPm mRNA in the absence of RFP fluorescence. Although there is no red fluorescence in cells transfected with pRFPm, RNA quantification shows there is no significant difference between RFP mRNA levels in cells transfected with pRFPwt, pRFPm, pRFPm- O 6 meG and pRFPm- O 6 meG plus 10 µM O 6 -benzylguanine. Additionally, treatment with 50 µM O 6 -benzylguanine significantly affected mRNA levels in cells transfected with pRFPm, and pRFPm- O 6 meG. Significance was determined by a two-sample Student’s t -test which compared the RFP–GFP mRNA abundance ratio from untreated pRwt transfected cells to the ratio observed under each of the experimental conditions in at least three independent experiments (* P

    Article Snippet: An aliquot of the digested DNA containing 0.05 ng (after DpnI digestion) or 0.008 ng (after DpnI and exonuclease III digestion) was used in a PCR reaction (GoTaq green master mix, Promega) containing primers (RdF: 5′-CAGCGGCCCTTCTCTCTTA-3′; RR: 5′-CGCTACAGGAACAGGTGGTG-3′) whose product spans 7 DpnI sites.

    Techniques: Polymerase Chain Reaction, Fluorescence, Transfection