top10 escherichia coli  (Thermo Fisher)


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    Structured Review

    Thermo Fisher top10 escherichia coli
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Top10 Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/top10 escherichia coli/product/Thermo Fisher
    Average 89 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    top10 escherichia coli - by Bioz Stars, 2020-05
    89/100 stars

    Images

    1) Product Images from "Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer"

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/cei.13183

    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Figure Legend Snippet: DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Techniques Used: DNA Methylation Assay, Sequencing, Cell Culture, Recombinant, In Vitro, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Expressing, Clone Assay, Plasmid Preparation, Transformation Assay

    2) Product Images from "Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer"

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/cei.13183

    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Figure Legend Snippet: DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Techniques Used: DNA Methylation Assay, Sequencing, Cell Culture, Recombinant, In Vitro, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Expressing, Clone Assay, Plasmid Preparation, Transformation Assay

    Related Articles

    Clone Assay:

    Article Title: Design and construction of generalizable RNA-protein hybrid controllers by level-matched genetic signal amplification
    Article Snippet: .. All cloned plasmid candidates were transformed into TOP10 Escherichia coli (Thermo Fisher Scientific), and plated on LB agar plates (EMD Millipore) with ampicillin (50 mg/L) for overnight growth in 37°C. .. The next day single colonies on the plates were first PCR screened with primers that bind 200~500 base pairs upstream and downstream of the inserts and further verified by sequencing.

    Article Title: Non-random pairing of CD46 isoforms with skewing towards BC2 and C2 in activated and memory/effector T cells
    Article Snippet: .. The plasmids were cloned in TOP10 E. coli (Invitrogen, USA) according to the manufacturer′s instructions. .. The orientation and correct sequence of the ORFs were verified by Sanger sequencing (GATC, Germany) using the following primers (TAG Copenhagen, Denmark) covering the insert: T7 fw 5′-TAATACGACTCACTATAGGG, and BGH rev: 5′-TAGAAGGCACAGTCGAGG.

    Article Title: Compensatory Mutations in Predicted Metal Transporters Modulate Auxin Conjugate Responsiveness in Arabidopsis
    Article Snippet: .. The MTP5 RT-PCR product was cloned into the pCR4Blunt-TOPO vector (Invitrogen, Carlsbad, CA) and transformed into TOP10 Escherichia coli (Invitrogen, Carlsbad, CA). .. Sequence analysis of the TOPO-MTP5 plasmid showed that the MTP5 sequence was mis-spliced (5′ of the mtp5-1 mutation) compared to annotation by The Arabidopsis Information Resource (TAIR; www.arabidopsis.org ).

    Amplification:

    Article Title: The Tyrosine Sulfate-rich Domains of the LRR Proteins Fibromodulin and Osteoadherin Bind Motifs of Basic Clusters in a Variety of Heparin-binding Proteins, Including Bioactive Factors *
    Article Snippet: .. The generated PCR fragment was ligated into the vector pET200/D-TOPO (Invitrogen) and amplified in TOP10 E. coli (Invitrogen). .. The sequence identity was confirmed, and the sequence was cloned into the expression vector pAM 104-2 and transformed into the Rosetta DE3TM E. coli (Novagen) expression system.

    Polymerase Chain Reaction:

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer
    Article Snippet: .. Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol. .. Transformed cells were cultured on lysogeny broth (LB) plates with 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) and incubated overnight at 37o C. The following day, 20 colonies were picked from each plate and cultured overnight in LB medium containing 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) at 37o C at 210 rpm.

    Article Title: The Tyrosine Sulfate-rich Domains of the LRR Proteins Fibromodulin and Osteoadherin Bind Motifs of Basic Clusters in a Variety of Heparin-binding Proteins, Including Bioactive Factors *
    Article Snippet: .. The generated PCR fragment was ligated into the vector pET200/D-TOPO (Invitrogen) and amplified in TOP10 E. coli (Invitrogen). .. The sequence identity was confirmed, and the sequence was cloned into the expression vector pAM 104-2 and transformed into the Rosetta DE3TM E. coli (Novagen) expression system.

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer
    Article Snippet: .. Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol. .. Transformed cells were cultured on lysogeny broth (LB) plates with 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) and incubated overnight at 37o C. The following day, 20 colonies were picked from each plate and cultured overnight in LB medium containing 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) at 37o C at 210 rpm.

    Generated:

    Article Title: The Tyrosine Sulfate-rich Domains of the LRR Proteins Fibromodulin and Osteoadherin Bind Motifs of Basic Clusters in a Variety of Heparin-binding Proteins, Including Bioactive Factors *
    Article Snippet: .. The generated PCR fragment was ligated into the vector pET200/D-TOPO (Invitrogen) and amplified in TOP10 E. coli (Invitrogen). .. The sequence identity was confirmed, and the sequence was cloned into the expression vector pAM 104-2 and transformed into the Rosetta DE3TM E. coli (Novagen) expression system.

    Expressing:

    Article Title: Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly
    Article Snippet: .. Growth and induction For all expression experiments in , vectors or isothermal assembly reactions were transformed into TOP10 E. coli (Invitrogen) using either the manufacturer's instructions or the TSS competent cell method as previously described ( ). .. Individual colonies were inoculated into 1 ml of LB + 1.0% glucose + 50 µg/ml ampicillin, and grown overnight at 30°C in a 2 ml deep-well plate (Thermo Scientific) with shaking at 1200 rpm on a Titramax 1000 platform shaker (Heidolph).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Compensatory Mutations in Predicted Metal Transporters Modulate Auxin Conjugate Responsiveness in Arabidopsis
    Article Snippet: .. The MTP5 RT-PCR product was cloned into the pCR4Blunt-TOPO vector (Invitrogen, Carlsbad, CA) and transformed into TOP10 Escherichia coli (Invitrogen, Carlsbad, CA). .. Sequence analysis of the TOPO-MTP5 plasmid showed that the MTP5 sequence was mis-spliced (5′ of the mtp5-1 mutation) compared to annotation by The Arabidopsis Information Resource (TAIR; www.arabidopsis.org ).

    Transformation Assay:

    Article Title: Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly
    Article Snippet: .. 2 µl of the assembly mixture was then transformed into TOP10 E. coli (Invitrogen). .. Growth and induction For all expression experiments in , vectors or isothermal assembly reactions were transformed into TOP10 E. coli (Invitrogen) using either the manufacturer's instructions or the TSS competent cell method as previously described ( ).

    Article Title: Design and construction of generalizable RNA-protein hybrid controllers by level-matched genetic signal amplification
    Article Snippet: .. All cloned plasmid candidates were transformed into TOP10 Escherichia coli (Thermo Fisher Scientific), and plated on LB agar plates (EMD Millipore) with ampicillin (50 mg/L) for overnight growth in 37°C. .. The next day single colonies on the plates were first PCR screened with primers that bind 200~500 base pairs upstream and downstream of the inserts and further verified by sequencing.

    Article Title: Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly
    Article Snippet: .. Growth and induction For all expression experiments in , vectors or isothermal assembly reactions were transformed into TOP10 E. coli (Invitrogen) using either the manufacturer's instructions or the TSS competent cell method as previously described ( ). .. Individual colonies were inoculated into 1 ml of LB + 1.0% glucose + 50 µg/ml ampicillin, and grown overnight at 30°C in a 2 ml deep-well plate (Thermo Scientific) with shaking at 1200 rpm on a Titramax 1000 platform shaker (Heidolph).

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer
    Article Snippet: .. Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol. .. Transformed cells were cultured on lysogeny broth (LB) plates with 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) and incubated overnight at 37o C. The following day, 20 colonies were picked from each plate and cultured overnight in LB medium containing 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) at 37o C at 210 rpm.

    Article Title: Compensatory Mutations in Predicted Metal Transporters Modulate Auxin Conjugate Responsiveness in Arabidopsis
    Article Snippet: .. The MTP5 RT-PCR product was cloned into the pCR4Blunt-TOPO vector (Invitrogen, Carlsbad, CA) and transformed into TOP10 Escherichia coli (Invitrogen, Carlsbad, CA). .. Sequence analysis of the TOPO-MTP5 plasmid showed that the MTP5 sequence was mis-spliced (5′ of the mtp5-1 mutation) compared to annotation by The Arabidopsis Information Resource (TAIR; www.arabidopsis.org ).

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer
    Article Snippet: .. Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol. .. Transformed cells were cultured on lysogeny broth (LB) plates with 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) and incubated overnight at 37o C. The following day, 20 colonies were picked from each plate and cultured overnight in LB medium containing 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) at 37o C at 210 rpm.

    Plasmid Preparation:

    Article Title: Design and construction of generalizable RNA-protein hybrid controllers by level-matched genetic signal amplification
    Article Snippet: .. All cloned plasmid candidates were transformed into TOP10 Escherichia coli (Thermo Fisher Scientific), and plated on LB agar plates (EMD Millipore) with ampicillin (50 mg/L) for overnight growth in 37°C. .. The next day single colonies on the plates were first PCR screened with primers that bind 200~500 base pairs upstream and downstream of the inserts and further verified by sequencing.

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer
    Article Snippet: .. Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol. .. Transformed cells were cultured on lysogeny broth (LB) plates with 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) and incubated overnight at 37o C. The following day, 20 colonies were picked from each plate and cultured overnight in LB medium containing 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) at 37o C at 210 rpm.

    Article Title: The Tyrosine Sulfate-rich Domains of the LRR Proteins Fibromodulin and Osteoadherin Bind Motifs of Basic Clusters in a Variety of Heparin-binding Proteins, Including Bioactive Factors *
    Article Snippet: .. The generated PCR fragment was ligated into the vector pET200/D-TOPO (Invitrogen) and amplified in TOP10 E. coli (Invitrogen). .. The sequence identity was confirmed, and the sequence was cloned into the expression vector pAM 104-2 and transformed into the Rosetta DE3TM E. coli (Novagen) expression system.

    Article Title: Compensatory Mutations in Predicted Metal Transporters Modulate Auxin Conjugate Responsiveness in Arabidopsis
    Article Snippet: .. The MTP5 RT-PCR product was cloned into the pCR4Blunt-TOPO vector (Invitrogen, Carlsbad, CA) and transformed into TOP10 Escherichia coli (Invitrogen, Carlsbad, CA). .. Sequence analysis of the TOPO-MTP5 plasmid showed that the MTP5 sequence was mis-spliced (5′ of the mtp5-1 mutation) compared to annotation by The Arabidopsis Information Resource (TAIR; www.arabidopsis.org ).

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer
    Article Snippet: .. Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol. .. Transformed cells were cultured on lysogeny broth (LB) plates with 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) and incubated overnight at 37o C. The following day, 20 colonies were picked from each plate and cultured overnight in LB medium containing 50 μg/ml Kanamycin (Substrate Unit, Karolinska University Hospital) at 37o C at 210 rpm.

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    Thermo Fisher top10 escherichia coli
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Top10 Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/top10 escherichia coli/product/Thermo Fisher
    Average 89 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    top10 escherichia coli - by Bioz Stars, 2020-05
    89/100 stars
      Buy from Supplier

    93
    Thermo Fisher e coli top10 carrying vector pcr4 topo
    Resistance against sulfonamide antibiotics mediated by SEW2_dhps01, SEW5_dhps01, AEW9_dhps01 , and AEG2_dhps01 . Five microliters of serially diluted E. coli <t>TOP10</t> cultures with starting OD 600 of 0.5 were spotted onto Iso-Sensitest agar plates supplemented with 1000 mg/L sulfamethazine (+ SMZ), 250 mg/L sulfamethoxazole (+ SMX), 250 mg/L sulfadiazine (+ SDZ) or 500 mg/L sulfisoxazole (+ SOX). Iso-Sensitest agar plates with no sulfonamide added (control) were also included. E. coli TOP10 cultures carrying the cloning vector <t>pCR4-TOPO,</t> pCR4_SEW2_dhps01, pCR4_SEW5_dhps01, pCR4_AEW9_dhps01 or pCR4_AEG2_dhps01 were considered.
    E Coli Top10 Carrying Vector Pcr4 Topo, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    92
    Thermo Fisher escherichia coli
    Representative SDS-PAGE, 2-dimensional electrophoresis (2DE), and Western blots showing the workflow for selection of immunoreactive spots. (A) The 1-dimensional SDS-PAGE image of Staphylococcus aureus Newbould 305 trypsinized protein (lane 1) and <t>Escherichia</t> coli trypsinized protein (lane 2). (B) 2DE gel conducted on 7 cm, pH 4 to 7, immobilized pH gradient (IPG) strips of S. aureus Newbould 305 trypsinized protein. Mr = molecular weight. (C) Immunoblot of S. aureus Newbould 305 trypsinized protein using pooled bovine mastitic milk, with immunoreactive regions highlighted in red circles. (D) 2DE gel conducted on 7 cm, pH 4 to 7, IPG strips of Escherichia coli DH10-β trypsinized protein. (E) Immunoblot of E. coli DH10-β trypsinized protein using bovine mastitic milk, with the immunoreactive regions highlighted in red circles. Color version available online.
    Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 389 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli/product/Thermo Fisher
    Average 92 stars, based on 389 article reviews
    Price from $9.99 to $1999.99
    escherichia coli - by Bioz Stars, 2020-05
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    Thermo Fisher e coli top10 cells
    Recombinant expression of rNanI species corresponding to protease-cleaved NanI fragments. rNanI species corresponding to full-length NanI or the major 65-kDa chymotrypsin-cleaved or 60-kDa mouse SI fluid-cleaved NanI fragments were recombinantly expressed by using E. coli <t>Top10</t> cells and the pTrc-HisB expression system. The rNanI species were then affinity enriched from the E. coli culture by using Talon resin and dialyzed overnight against PBS. Panel A shows the dialyzed samples subjected to SDS-PAGE and then either stained with Coomassie blue (left) or Western blotted for NanI (right). The relative amount of each enriched rNanI species was then determined by a protein assay. All experiments were performed in triplicate, and the mean results are shown. Error bars show standard deviations. # indicates a significant ( P
    E Coli Top10 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 7 article reviews
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    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Journal: Clinical and Experimental Immunology

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer

    doi: 10.1111/cei.13183

    Figure Lengend Snippet: DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Article Snippet: Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol.

    Techniques: DNA Methylation Assay, Sequencing, Cell Culture, Recombinant, In Vitro, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Expressing, Clone Assay, Plasmid Preparation, Transformation Assay

    Resistance against sulfonamide antibiotics mediated by SEW2_dhps01, SEW5_dhps01, AEW9_dhps01 , and AEG2_dhps01 . Five microliters of serially diluted E. coli TOP10 cultures with starting OD 600 of 0.5 were spotted onto Iso-Sensitest agar plates supplemented with 1000 mg/L sulfamethazine (+ SMZ), 250 mg/L sulfamethoxazole (+ SMX), 250 mg/L sulfadiazine (+ SDZ) or 500 mg/L sulfisoxazole (+ SOX). Iso-Sensitest agar plates with no sulfonamide added (control) were also included. E. coli TOP10 cultures carrying the cloning vector pCR4-TOPO, pCR4_SEW2_dhps01, pCR4_SEW5_dhps01, pCR4_AEW9_dhps01 or pCR4_AEG2_dhps01 were considered.

    Journal: Frontiers in Microbiology

    Article Title: Discovery of Novel Antibiotic Resistance Determinants in Forest and Grassland Soil Metagenomes

    doi: 10.3389/fmicb.2019.00460

    Figure Lengend Snippet: Resistance against sulfonamide antibiotics mediated by SEW2_dhps01, SEW5_dhps01, AEW9_dhps01 , and AEG2_dhps01 . Five microliters of serially diluted E. coli TOP10 cultures with starting OD 600 of 0.5 were spotted onto Iso-Sensitest agar plates supplemented with 1000 mg/L sulfamethazine (+ SMZ), 250 mg/L sulfamethoxazole (+ SMX), 250 mg/L sulfadiazine (+ SDZ) or 500 mg/L sulfisoxazole (+ SOX). Iso-Sensitest agar plates with no sulfonamide added (control) were also included. E. coli TOP10 cultures carrying the cloning vector pCR4-TOPO, pCR4_SEW2_dhps01, pCR4_SEW5_dhps01, pCR4_AEW9_dhps01 or pCR4_AEG2_dhps01 were considered.

    Article Snippet: E. coli TOP10 carrying vector pCR4-TOPO (Thermo Fisher Scientific) was used as control.

    Techniques: Clone Assay, Plasmid Preparation

    Antibiotic susceptibility profiles of E. coli TOP10 carrying soil-derived genes involved in antibiotic resistance. The genes were subcloned into plasmid vector pCR4-TOPO. MICs of antibiotics were determined using the broth microdilution method and are presented as fold increase relative to those for E. coli TOP10 carrying the cloning vector pCR4-TOPO. CAX, cefotaxime; CHL, chloramphenicol; ERY, erythromycin; GEN, gentamicin; LIN, lincomycin; RIF, rifampicin; SDZ, sulfadiazine; SMX, sulfamethoxazole; SMZ, sulfamethazine; SOX, sulfisoxazole; TET, tetracycline; TYL, tylosin.

    Journal: Frontiers in Microbiology

    Article Title: Discovery of Novel Antibiotic Resistance Determinants in Forest and Grassland Soil Metagenomes

    doi: 10.3389/fmicb.2019.00460

    Figure Lengend Snippet: Antibiotic susceptibility profiles of E. coli TOP10 carrying soil-derived genes involved in antibiotic resistance. The genes were subcloned into plasmid vector pCR4-TOPO. MICs of antibiotics were determined using the broth microdilution method and are presented as fold increase relative to those for E. coli TOP10 carrying the cloning vector pCR4-TOPO. CAX, cefotaxime; CHL, chloramphenicol; ERY, erythromycin; GEN, gentamicin; LIN, lincomycin; RIF, rifampicin; SDZ, sulfadiazine; SMX, sulfamethoxazole; SMZ, sulfamethazine; SOX, sulfisoxazole; TET, tetracycline; TYL, tylosin.

    Article Snippet: E. coli TOP10 carrying vector pCR4-TOPO (Thermo Fisher Scientific) was used as control.

    Techniques: Derivative Assay, Plasmid Preparation, Clone Assay

    Representative SDS-PAGE, 2-dimensional electrophoresis (2DE), and Western blots showing the workflow for selection of immunoreactive spots. (A) The 1-dimensional SDS-PAGE image of Staphylococcus aureus Newbould 305 trypsinized protein (lane 1) and Escherichia coli trypsinized protein (lane 2). (B) 2DE gel conducted on 7 cm, pH 4 to 7, immobilized pH gradient (IPG) strips of S. aureus Newbould 305 trypsinized protein. Mr = molecular weight. (C) Immunoblot of S. aureus Newbould 305 trypsinized protein using pooled bovine mastitic milk, with immunoreactive regions highlighted in red circles. (D) 2DE gel conducted on 7 cm, pH 4 to 7, IPG strips of Escherichia coli DH10-β trypsinized protein. (E) Immunoblot of E. coli DH10-β trypsinized protein using bovine mastitic milk, with the immunoreactive regions highlighted in red circles. Color version available online.

    Journal: Journal of dairy science

    Article Title: Immunoproteomics to identify Staphylococcus aureus antigens expressed in bovine milk during mastitis

    doi: 10.3168/jds.2017-14040

    Figure Lengend Snippet: Representative SDS-PAGE, 2-dimensional electrophoresis (2DE), and Western blots showing the workflow for selection of immunoreactive spots. (A) The 1-dimensional SDS-PAGE image of Staphylococcus aureus Newbould 305 trypsinized protein (lane 1) and Escherichia coli trypsinized protein (lane 2). (B) 2DE gel conducted on 7 cm, pH 4 to 7, immobilized pH gradient (IPG) strips of S. aureus Newbould 305 trypsinized protein. Mr = molecular weight. (C) Immunoblot of S. aureus Newbould 305 trypsinized protein using pooled bovine mastitic milk, with immunoreactive regions highlighted in red circles. (D) 2DE gel conducted on 7 cm, pH 4 to 7, IPG strips of Escherichia coli DH10-β trypsinized protein. (E) Immunoblot of E. coli DH10-β trypsinized protein using bovine mastitic milk, with the immunoreactive regions highlighted in red circles. Color version available online.

    Article Snippet: Briefly, S. aureus Newbould 305, S. aureus C1, and Escherichia coli (DH10-β Top10, Thermo Fisher Scientific, Waltham, MA) were grown in LIM overnight to an optical density ( OD ) of 0.75 to 1.2 and harvested by centrifugation (6,000 × g for 10 min at 4°C), before washing 3 times with 1× PBS.

    Techniques: SDS Page, Electrophoresis, Two-Dimensional Gel Electrophoresis, Western Blot, Selection, Molecular Weight

    Recombinant expression of rNanI species corresponding to protease-cleaved NanI fragments. rNanI species corresponding to full-length NanI or the major 65-kDa chymotrypsin-cleaved or 60-kDa mouse SI fluid-cleaved NanI fragments were recombinantly expressed by using E. coli Top10 cells and the pTrc-HisB expression system. The rNanI species were then affinity enriched from the E. coli culture by using Talon resin and dialyzed overnight against PBS. Panel A shows the dialyzed samples subjected to SDS-PAGE and then either stained with Coomassie blue (left) or Western blotted for NanI (right). The relative amount of each enriched rNanI species was then determined by a protein assay. All experiments were performed in triplicate, and the mean results are shown. Error bars show standard deviations. # indicates a significant ( P

    Journal: Infection and Immunity

    Article Title: Native or Proteolytically Activated NanI Sialidase Enhances the Binding and Cytotoxic Activity of Clostridium perfringens Enterotoxin and Beta Toxin

    doi: 10.1128/IAI.00730-17

    Figure Lengend Snippet: Recombinant expression of rNanI species corresponding to protease-cleaved NanI fragments. rNanI species corresponding to full-length NanI or the major 65-kDa chymotrypsin-cleaved or 60-kDa mouse SI fluid-cleaved NanI fragments were recombinantly expressed by using E. coli Top10 cells and the pTrc-HisB expression system. The rNanI species were then affinity enriched from the E. coli culture by using Talon resin and dialyzed overnight against PBS. Panel A shows the dialyzed samples subjected to SDS-PAGE and then either stained with Coomassie blue (left) or Western blotted for NanI (right). The relative amount of each enriched rNanI species was then determined by a protein assay. All experiments were performed in triplicate, and the mean results are shown. Error bars show standard deviations. # indicates a significant ( P

    Article Snippet: E. coli Top10 cells (ThermoFisher) were routinely grown on LB medium (Fisher Scientific) supplemented with 100 μg/ml of ampicillin (Fisher Scientific), as indicated.

    Techniques: Recombinant, Expressing, SDS Page, Staining, Western Blot