enhancer solution  (Thermo Fisher)


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    Name:
    Enhancer Solution
    Description:
    The InstantOne ELISA Enhancer Solution is a supplement added to the 5X Cell Lysis Buffer Cat No IOCLB1 to yield the Cell Lysis Mix solution that is applied to cultured cells or tissues prior to running the assays The 1X Cell Lysis Mix is prepared by simply diluting Cell Lysis Mix 5X a mixture of the Cell Lysis Buffer 5X and the Enhancer Solution 5 fold with deionized water This buffer is used to lyse cells after the removal of culture medium and is typically used to lyse adherent cells or non adherent cells that have been harvested by centrifugation Lysis Mix should be used as the diluent for any dilution of cellular lysates that are required Reported ApplicationELISA
    Catalog Number:
    ioes1
    Price:
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    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher enhancer solution
    The InstantOne ELISA Enhancer Solution is a supplement added to the 5X Cell Lysis Buffer Cat No IOCLB1 to yield the Cell Lysis Mix solution that is applied to cultured cells or tissues prior to running the assays The 1X Cell Lysis Mix is prepared by simply diluting Cell Lysis Mix 5X a mixture of the Cell Lysis Buffer 5X and the Enhancer Solution 5 fold with deionized water This buffer is used to lyse cells after the removal of culture medium and is typically used to lyse adherent cells or non adherent cells that have been harvested by centrifugation Lysis Mix should be used as the diluent for any dilution of cellular lysates that are required Reported ApplicationELISA
    https://www.bioz.com/result/enhancer solution/product/Thermo Fisher
    Average 99 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    enhancer solution - by Bioz Stars, 2020-07
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Defective Erythropoiesis in Transgenic Mice Expressing Dominant-Negative Upstream Stimulatory Factor ▿
    Article Snippet: .. The 3′ enhancer and 3′ chicken HS4 elements were amplified using pITRp543f2beta4 as the template with primers containing PmeI sites and were cloned into the pTOPO vector (Invitrogen). .. The pTOPO/3′ enhancer, 3′ chicken HS4, and pITRp543f2A-USF subsequently were digested with PmeI, and the appropriate fragments were isolated and ligated to form the vector pITRp543f2A-USF4.

    Amplification:

    Article Title: Population Structure of Bartonella henselae in Algerian Urban Stray Cats
    Article Snippet: .. Briefly, each BHV PCR mix contained 5 µl cell lysate, 2.5 µl 10× Pfx amplification buffer, 1 µl of 10 mM DNTP mixture, 0.5 µl MgSO4 (50 mM), 2 µl of each reverse and forward primers (10µM), 2.5 µl of 10X PCR enhancer solution and 0.4 µl platinum Pfx DNA polymerase (2.5 units/µl) (InVitrogen, California, USA). .. PCR amplification program consisted of a denaturation step at 94°C for 5 mn followed by 34 amplification cycles: 94°C for 30 s, 53°C for 30 s and 72°C for 2 mn, with a final extension step of 10 mn at 72°C.

    Article Title: Defective Erythropoiesis in Transgenic Mice Expressing Dominant-Negative Upstream Stimulatory Factor ▿
    Article Snippet: .. The 3′ enhancer and 3′ chicken HS4 elements were amplified using pITRp543f2beta4 as the template with primers containing PmeI sites and were cloned into the pTOPO vector (Invitrogen). .. The pTOPO/3′ enhancer, 3′ chicken HS4, and pITRp543f2A-USF subsequently were digested with PmeI, and the appropriate fragments were isolated and ligated to form the vector pITRp543f2A-USF4.

    Article Title: Predominant Clonal Accumulation of CD8+ T Cells with Moderate Avidity in the Central Nervous Systems of Theiler's Virus-Infected C57BL/6 Mice ▿
    Article Snippet: .. Primary amplification of cDNAs from isolated antigen-specific T cells was performed for 35 cycles using Vβ-Cβ primer pairs at 58°C for annealing in the presence of PCR enhancer solution (GIBCO, Invitrogen Corporation, Grand Island, NY). .. The primary PCR product (1 μl) was subjected to a secondary PCR with a 32 P-end-labeled Vβ chain primer and 12 different Jβ primers (Table ).

    Article Title: FRA2A Is a CGG Repeat Expansion Associated with Silencing of AFF3
    Article Snippet: .. PCR amplification and hybridization of the FRA2A-associated CGG repeat PCR amplification of the normal FRA2A CGG repeat was performed with the aid of 2.5× PCR Enhancer solution (Invitrogen, Carlsbad, CA, USA) using a forward primer P1 ( 5′-GGCCGTAAAAGCCACGAGAGAGGG-3′ ) and a reverse primer P2 ( 5′-CTTGCGCGCAGGCACACTCAAGAG-3′ ) derived from the sequences flanking the repeat. .. PCR products were sequenced and subsequently analysed by use of an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

    Isolation:

    Article Title: Predominant Clonal Accumulation of CD8+ T Cells with Moderate Avidity in the Central Nervous Systems of Theiler's Virus-Infected C57BL/6 Mice ▿
    Article Snippet: .. Primary amplification of cDNAs from isolated antigen-specific T cells was performed for 35 cycles using Vβ-Cβ primer pairs at 58°C for annealing in the presence of PCR enhancer solution (GIBCO, Invitrogen Corporation, Grand Island, NY). .. The primary PCR product (1 μl) was subjected to a secondary PCR with a 32 P-end-labeled Vβ chain primer and 12 different Jβ primers (Table ).

    Derivative Assay:

    Article Title: FRA2A Is a CGG Repeat Expansion Associated with Silencing of AFF3
    Article Snippet: .. PCR amplification and hybridization of the FRA2A-associated CGG repeat PCR amplification of the normal FRA2A CGG repeat was performed with the aid of 2.5× PCR Enhancer solution (Invitrogen, Carlsbad, CA, USA) using a forward primer P1 ( 5′-GGCCGTAAAAGCCACGAGAGAGGG-3′ ) and a reverse primer P2 ( 5′-CTTGCGCGCAGGCACACTCAAGAG-3′ ) derived from the sequences flanking the repeat. .. PCR products were sequenced and subsequently analysed by use of an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

    Polymerase Chain Reaction:

    Article Title: Population Structure of Bartonella henselae in Algerian Urban Stray Cats
    Article Snippet: .. Briefly, each BHV PCR mix contained 5 µl cell lysate, 2.5 µl 10× Pfx amplification buffer, 1 µl of 10 mM DNTP mixture, 0.5 µl MgSO4 (50 mM), 2 µl of each reverse and forward primers (10µM), 2.5 µl of 10X PCR enhancer solution and 0.4 µl platinum Pfx DNA polymerase (2.5 units/µl) (InVitrogen, California, USA). .. PCR amplification program consisted of a denaturation step at 94°C for 5 mn followed by 34 amplification cycles: 94°C for 30 s, 53°C for 30 s and 72°C for 2 mn, with a final extension step of 10 mn at 72°C.

    Article Title: Predominant Clonal Accumulation of CD8+ T Cells with Moderate Avidity in the Central Nervous Systems of Theiler's Virus-Infected C57BL/6 Mice ▿
    Article Snippet: .. Primary amplification of cDNAs from isolated antigen-specific T cells was performed for 35 cycles using Vβ-Cβ primer pairs at 58°C for annealing in the presence of PCR enhancer solution (GIBCO, Invitrogen Corporation, Grand Island, NY). .. The primary PCR product (1 μl) was subjected to a secondary PCR with a 32 P-end-labeled Vβ chain primer and 12 different Jβ primers (Table ).

    Article Title: FRA2A Is a CGG Repeat Expansion Associated with Silencing of AFF3
    Article Snippet: .. PCR amplification and hybridization of the FRA2A-associated CGG repeat PCR amplification of the normal FRA2A CGG repeat was performed with the aid of 2.5× PCR Enhancer solution (Invitrogen, Carlsbad, CA, USA) using a forward primer P1 ( 5′-GGCCGTAAAAGCCACGAGAGAGGG-3′ ) and a reverse primer P2 ( 5′-CTTGCGCGCAGGCACACTCAAGAG-3′ ) derived from the sequences flanking the repeat. .. PCR products were sequenced and subsequently analysed by use of an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

    Plasmid Preparation:

    Article Title: Defective Erythropoiesis in Transgenic Mice Expressing Dominant-Negative Upstream Stimulatory Factor ▿
    Article Snippet: .. The 3′ enhancer and 3′ chicken HS4 elements were amplified using pITRp543f2beta4 as the template with primers containing PmeI sites and were cloned into the pTOPO vector (Invitrogen). .. The pTOPO/3′ enhancer, 3′ chicken HS4, and pITRp543f2A-USF subsequently were digested with PmeI, and the appropriate fragments were isolated and ligated to form the vector pITRp543f2A-USF4.

    Hybridization:

    Article Title: FRA2A Is a CGG Repeat Expansion Associated with Silencing of AFF3
    Article Snippet: .. PCR amplification and hybridization of the FRA2A-associated CGG repeat PCR amplification of the normal FRA2A CGG repeat was performed with the aid of 2.5× PCR Enhancer solution (Invitrogen, Carlsbad, CA, USA) using a forward primer P1 ( 5′-GGCCGTAAAAGCCACGAGAGAGGG-3′ ) and a reverse primer P2 ( 5′-CTTGCGCGCAGGCACACTCAAGAG-3′ ) derived from the sequences flanking the repeat. .. PCR products were sequenced and subsequently analysed by use of an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

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  • 99
    Thermo Fisher luminol enhancer solution
    EtpE-C-coated beads block PMA-induced ROS generation by WT BMDM but not CD147 –/– BMDM. BMDM from WT mice (A and B) or CD147 –/– mice (C and D) were preincubated with <t>luminol</t> in HBSSd for 15 min and then incubated at 37°C for 30 min with beads coated with 40 ng of EtpE-C or EtpE-N or with uncoated beads or with HBSSd (control [CTL]). ROS generation was induced with PMA, recorded (A and C), and analyzed (B and D), and the results are presented as in Fig. 2 . UN, unstimulated BMDM in HBSSd without PMA addition. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P
    Luminol Enhancer Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/luminol enhancer solution/product/Thermo Fisher
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    luminol enhancer solution - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    88
    Thermo Fisher metal enhanced diaminobenzidine dab solution
    EtpE-C-coated beads block PMA-induced ROS generation by WT BMDM but not CD147 –/– BMDM. BMDM from WT mice (A and B) or CD147 –/– mice (C and D) were preincubated with <t>luminol</t> in HBSSd for 15 min and then incubated at 37°C for 30 min with beads coated with 40 ng of EtpE-C or EtpE-N or with uncoated beads or with HBSSd (control [CTL]). ROS generation was induced with PMA, recorded (A and C), and analyzed (B and D), and the results are presented as in Fig. 2 . UN, unstimulated BMDM in HBSSd without PMA addition. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P
    Metal Enhanced Diaminobenzidine Dab Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/metal enhanced diaminobenzidine dab solution/product/Thermo Fisher
    Average 88 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    metal enhanced diaminobenzidine dab solution - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    Image Search Results


    EtpE-C-coated beads block PMA-induced ROS generation by WT BMDM but not CD147 –/– BMDM. BMDM from WT mice (A and B) or CD147 –/– mice (C and D) were preincubated with luminol in HBSSd for 15 min and then incubated at 37°C for 30 min with beads coated with 40 ng of EtpE-C or EtpE-N or with uncoated beads or with HBSSd (control [CTL]). ROS generation was induced with PMA, recorded (A and C), and analyzed (B and D), and the results are presented as in Fig. 2 . UN, unstimulated BMDM in HBSSd without PMA addition. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis Uses an Invasin To Suppress Reactive Oxygen Species Generation by Macrophages via CD147-Dependent Inhibition of Vav1 To Block Rac1 Activation

    doi: 10.1128/mBio.00267-20

    Figure Lengend Snippet: EtpE-C-coated beads block PMA-induced ROS generation by WT BMDM but not CD147 –/– BMDM. BMDM from WT mice (A and B) or CD147 –/– mice (C and D) were preincubated with luminol in HBSSd for 15 min and then incubated at 37°C for 30 min with beads coated with 40 ng of EtpE-C or EtpE-N or with uncoated beads or with HBSSd (control [CTL]). ROS generation was induced with PMA, recorded (A and C), and analyzed (B and D), and the results are presented as in Fig. 2 . UN, unstimulated BMDM in HBSSd without PMA addition. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P

    Article Snippet: Immunopositive bands were detected with an enhanced chemiluminescence detection method using peroxide solution (Thermo Fisher Scientific, Rockford, IL) and luminol enhancer solution (Thermo Fisher Scientific).

    Techniques: Blocking Assay, Mouse Assay, Incubation

    Nucleofection with CD147, but not control GFP plasmid, complements BMDM from CD147 –/– mice for suppression of ROS generation by E. chaffeensis . BMDM from CD147 –/– mice were nucleofected with a CD147-encoding plasmid (CD147) or control plasmid for 1 day (A). Nucleofected cells were preincubated with luminol in HBSSd for 15 min and then incubated with E. chaffeensis ( Ech ), DH82 cell lysate, or HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was induced with PMA, recorded (A and C), and analyzed (B and D). UN, unstimulated BMDM in HBSSd without PMA addition. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis Uses an Invasin To Suppress Reactive Oxygen Species Generation by Macrophages via CD147-Dependent Inhibition of Vav1 To Block Rac1 Activation

    doi: 10.1128/mBio.00267-20

    Figure Lengend Snippet: Nucleofection with CD147, but not control GFP plasmid, complements BMDM from CD147 –/– mice for suppression of ROS generation by E. chaffeensis . BMDM from CD147 –/– mice were nucleofected with a CD147-encoding plasmid (CD147) or control plasmid for 1 day (A). Nucleofected cells were preincubated with luminol in HBSSd for 15 min and then incubated with E. chaffeensis ( Ech ), DH82 cell lysate, or HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was induced with PMA, recorded (A and C), and analyzed (B and D). UN, unstimulated BMDM in HBSSd without PMA addition. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P

    Article Snippet: Immunopositive bands were detected with an enhanced chemiluminescence detection method using peroxide solution (Thermo Fisher Scientific, Rockford, IL) and luminol enhancer solution (Thermo Fisher Scientific).

    Techniques: Plasmid Preparation, Mouse Assay, Incubation

    EtpE-C-coated beads do not block PMA-induced ROS generation in human monocytes pretreated with anti-CD147. Human monocytes were preincubated with luminol with or without 10 μg/ml anti-CD147 (α-CD147) for 30 min at 37°C and then incubated with beads coated with 40 ng of EtpE-C or EtpE-N (∼5 × 10 6 beads) or with HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was continuously recorded as the relative chemiluminescence of oxidized luminol after the addition of PMA (200 nM, indicated by arrows) (A). UN, unstimulated human monocytes in HBSSd without PMA addition. The area under the curve was measured over 30 min after PMA addition and is shown relative to ROS generation in the control with PMA, which was considered 100% in panel B. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis Uses an Invasin To Suppress Reactive Oxygen Species Generation by Macrophages via CD147-Dependent Inhibition of Vav1 To Block Rac1 Activation

    doi: 10.1128/mBio.00267-20

    Figure Lengend Snippet: EtpE-C-coated beads do not block PMA-induced ROS generation in human monocytes pretreated with anti-CD147. Human monocytes were preincubated with luminol with or without 10 μg/ml anti-CD147 (α-CD147) for 30 min at 37°C and then incubated with beads coated with 40 ng of EtpE-C or EtpE-N (∼5 × 10 6 beads) or with HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was continuously recorded as the relative chemiluminescence of oxidized luminol after the addition of PMA (200 nM, indicated by arrows) (A). UN, unstimulated human monocytes in HBSSd without PMA addition. The area under the curve was measured over 30 min after PMA addition and is shown relative to ROS generation in the control with PMA, which was considered 100% in panel B. Results represent the means plus SD from at least three independent experiments and were compared by Student’s t test. * , P

    Article Snippet: Immunopositive bands were detected with an enhanced chemiluminescence detection method using peroxide solution (Thermo Fisher Scientific, Rockford, IL) and luminol enhancer solution (Thermo Fisher Scientific).

    Techniques: Blocking Assay, Incubation

    E. chaffeensis blocks PMA-induced ROS generation by WT BMDM but not by CD147 –/– BMDM. BMDM from WT mice (A and B) or CD147 –/– mice (C and D) were preincubated with luminol in HBSSd for 15 min and then incubated with E. chaffeensis ( Ech ) isolated from infected DH82 cells, DH82 cell lysate, or HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was continuously recorded as the relative chemiluminescence of oxidized luminol after the addition of PMA (200 nM, indicated by arrows) (A and C). UN, unstimulated BMDM in HBSSd without PMA addition. The area under the curve was measured over 60 min after PMA addition and is shown relative to ROS generation in the control with PMA, which was considered 100% in panels B and D. Results represent the means plus standard deviations (SD) (error bars) from at least three independent experiments and were compared by Student’s t test. Values that are significantly different ( P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis Uses an Invasin To Suppress Reactive Oxygen Species Generation by Macrophages via CD147-Dependent Inhibition of Vav1 To Block Rac1 Activation

    doi: 10.1128/mBio.00267-20

    Figure Lengend Snippet: E. chaffeensis blocks PMA-induced ROS generation by WT BMDM but not by CD147 –/– BMDM. BMDM from WT mice (A and B) or CD147 –/– mice (C and D) were preincubated with luminol in HBSSd for 15 min and then incubated with E. chaffeensis ( Ech ) isolated from infected DH82 cells, DH82 cell lysate, or HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was continuously recorded as the relative chemiluminescence of oxidized luminol after the addition of PMA (200 nM, indicated by arrows) (A and C). UN, unstimulated BMDM in HBSSd without PMA addition. The area under the curve was measured over 60 min after PMA addition and is shown relative to ROS generation in the control with PMA, which was considered 100% in panels B and D. Results represent the means plus standard deviations (SD) (error bars) from at least three independent experiments and were compared by Student’s t test. Values that are significantly different ( P

    Article Snippet: Immunopositive bands were detected with an enhanced chemiluminescence detection method using peroxide solution (Thermo Fisher Scientific, Rockford, IL) and luminol enhancer solution (Thermo Fisher Scientific).

    Techniques: Mouse Assay, Incubation, Isolation, Infection

    Recombinant gallin identified by western blotting performed using a SuperSignal ® West HisProbe™ Kit (Pierce, Rockford IL and visualised with Luminol/Enhancer solution with stable Peroxide Solution (Thermo, USA) . The membrane was exposed to X-ray film for 30 seconds. The lanes show the signal from samples derived from either the supernatant or the bacterial pellet after induction by 1 mM IPTG for 0, 2, 3 5 or 15 hours. Molecular weight was determined by interpolation from a Spectra™ Multicolor Broad Range Protein Ladder (Fermentas) run in lane M.

    Journal: BMC Immunology

    Article Title: Gallin; an antimicrobial peptide member of a new avian defensin family, the ovodefensins, has been subject to recent gene duplication

    doi: 10.1186/1471-2172-11-12

    Figure Lengend Snippet: Recombinant gallin identified by western blotting performed using a SuperSignal ® West HisProbe™ Kit (Pierce, Rockford IL and visualised with Luminol/Enhancer solution with stable Peroxide Solution (Thermo, USA) . The membrane was exposed to X-ray film for 30 seconds. The lanes show the signal from samples derived from either the supernatant or the bacterial pellet after induction by 1 mM IPTG for 0, 2, 3 5 or 15 hours. Molecular weight was determined by interpolation from a Spectra™ Multicolor Broad Range Protein Ladder (Fermentas) run in lane M.

    Article Snippet: Finally the chemiluminescence detection reaction was performed by using equal volumes of Luminol/Enhancer solution with stable Peroxide Solution (Thermo, USA), and the membrane was exposed to X-ray film for 30 seconds.

    Techniques: Recombinant, Western Blot, Derivative Assay, Molecular Weight

    EtpE-C prevents latex bead-induced ROS generation by BMDMs from WT, but not DNase X −/− , mice. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with 40 ng of EtpE-C- or EtpE-N-coated or uncoated beads (~5 × 10 6 ) or HBSSd (control [CTL]), and ROS generation was recorded as the relative chemiluminescence of oxidized luminol (A, C). The area under the curve was measured over 30 min after bead addition and is shown relative to ROS generation with uncoated beads, which was considered 100% (B, D). Results are presented as the mean ± the standard deviation of at least three independent experiments and were compared with a Student t test; *, P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis and Its Invasin EtpE Block Reactive Oxygen Species Generation by Macrophages in a DNase X-Dependent Manner

    doi: 10.1128/mBio.01551-17

    Figure Lengend Snippet: EtpE-C prevents latex bead-induced ROS generation by BMDMs from WT, but not DNase X −/− , mice. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with 40 ng of EtpE-C- or EtpE-N-coated or uncoated beads (~5 × 10 6 ) or HBSSd (control [CTL]), and ROS generation was recorded as the relative chemiluminescence of oxidized luminol (A, C). The area under the curve was measured over 30 min after bead addition and is shown relative to ROS generation with uncoated beads, which was considered 100% (B, D). Results are presented as the mean ± the standard deviation of at least three independent experiments and were compared with a Student t test; *, P

    Article Snippet: Reactive bands were visualized by enhanced chemiluminescence by incubating the membrane with the respective horseradish peroxidase-conjugated secondary antibodies and the same volume of H2 O2 solution (Thermo Scientific) and Luminol/Enhancer solution (Thermo Scientific).

    Techniques: Mouse Assay, Incubation, CTL Assay, Standard Deviation

    EtpE-C prevents latex bead-induced ROS generation by BMDMs from WT mice in a dose-dependent manner. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with beads coated with 40, 20, or 0 ng of EtpE-C (~5 × 10 6 ) or with HBSSd (control [CTL]), and ROS generation was recorded as the relative chemiluminescence of oxidized luminol (A, C). The area under the curve was measured over 60 min after bead addition and is shown relative to ROS generation with uncoated (0 ng of EtpE-C) beads, which was considered 100% (B, D). Results are presented as the mean ± the standard deviation of at least three independent experiments. The coefficient of correlation ( r value) between the relative levels of ROS and the amounts of EtpE-C is 0.999 ( P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis and Its Invasin EtpE Block Reactive Oxygen Species Generation by Macrophages in a DNase X-Dependent Manner

    doi: 10.1128/mBio.01551-17

    Figure Lengend Snippet: EtpE-C prevents latex bead-induced ROS generation by BMDMs from WT mice in a dose-dependent manner. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with beads coated with 40, 20, or 0 ng of EtpE-C (~5 × 10 6 ) or with HBSSd (control [CTL]), and ROS generation was recorded as the relative chemiluminescence of oxidized luminol (A, C). The area under the curve was measured over 60 min after bead addition and is shown relative to ROS generation with uncoated (0 ng of EtpE-C) beads, which was considered 100% (B, D). Results are presented as the mean ± the standard deviation of at least three independent experiments. The coefficient of correlation ( r value) between the relative levels of ROS and the amounts of EtpE-C is 0.999 ( P

    Article Snippet: Reactive bands were visualized by enhanced chemiluminescence by incubating the membrane with the respective horseradish peroxidase-conjugated secondary antibodies and the same volume of H2 O2 solution (Thermo Scientific) and Luminol/Enhancer solution (Thermo Scientific).

    Techniques: Mouse Assay, Incubation, CTL Assay, Standard Deviation

    EtpE-C-coated beads block PMA-induced ROS generation by BMDMs from WT mice but not by BMDMs from DNase X −/− mice. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with beads coated with 40 ng of EtpE-C or EtpE-N, uncoated beads (~5 × 10 6 ), or with HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was induced with PMA, recorded (A, C), and analyzed (B, D), and the results are presented as in Fig. 1 ; UN, unstimulated BMDMs in HBSSd without PMA addition. *, P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis and Its Invasin EtpE Block Reactive Oxygen Species Generation by Macrophages in a DNase X-Dependent Manner

    doi: 10.1128/mBio.01551-17

    Figure Lengend Snippet: EtpE-C-coated beads block PMA-induced ROS generation by BMDMs from WT mice but not by BMDMs from DNase X −/− mice. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with beads coated with 40 ng of EtpE-C or EtpE-N, uncoated beads (~5 × 10 6 ), or with HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was induced with PMA, recorded (A, C), and analyzed (B, D), and the results are presented as in Fig. 1 ; UN, unstimulated BMDMs in HBSSd without PMA addition. *, P

    Article Snippet: Reactive bands were visualized by enhanced chemiluminescence by incubating the membrane with the respective horseradish peroxidase-conjugated secondary antibodies and the same volume of H2 O2 solution (Thermo Scientific) and Luminol/Enhancer solution (Thermo Scientific).

    Techniques: Blocking Assay, Mouse Assay, Incubation, CTL Assay

    Entry of rEtpE-C-coated beads into WT BMDMs is not required for inhibition of ROS generation in response to PMA. BMDMs from WT mice were preincubated with luminol with or without 10 µM wiskostatin for 30 min at 37°C and then incubated with beads coated with 40 ng of EtpE-C or EtpE-N (~5 × 10 6 beads) or with HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was induced with PMA, recorded (A), and analyzed (B), and the results are presented as in Fig. 1 ; UN, unstimulated BMDMs in HBSSd without PMA addition. *, P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis and Its Invasin EtpE Block Reactive Oxygen Species Generation by Macrophages in a DNase X-Dependent Manner

    doi: 10.1128/mBio.01551-17

    Figure Lengend Snippet: Entry of rEtpE-C-coated beads into WT BMDMs is not required for inhibition of ROS generation in response to PMA. BMDMs from WT mice were preincubated with luminol with or without 10 µM wiskostatin for 30 min at 37°C and then incubated with beads coated with 40 ng of EtpE-C or EtpE-N (~5 × 10 6 beads) or with HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was induced with PMA, recorded (A), and analyzed (B), and the results are presented as in Fig. 1 ; UN, unstimulated BMDMs in HBSSd without PMA addition. *, P

    Article Snippet: Reactive bands were visualized by enhanced chemiluminescence by incubating the membrane with the respective horseradish peroxidase-conjugated secondary antibodies and the same volume of H2 O2 solution (Thermo Scientific) and Luminol/Enhancer solution (Thermo Scientific).

    Techniques: Inhibition, Mouse Assay, Incubation, CTL Assay

    E. chaffeensis blocks PMA-induced ROS generation by BMDMs from WT mice but not by BMDMs from DNase X −/− mice. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with E. chaffeensis ( Ech ) isolated from infected DH82 cells, DH82 cell lysate, or HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was continuously recorded as the relative chemiluminescence of oxidized luminol after the addition of PMA (0.5 µg/ml, indicated by arrows) (A, C). UN, unstimulated BMDMs in HBSSd without PMA addition. The area under the curve was measured over 60 min after PMA addition and is shown relative to ROS generation in the control with PMA, which was considered 100% (B, D). Results are presented as the mean ± the standard deviation from at least three independent experiments and were compared with a Student t test; *, P

    Journal: mBio

    Article Title: Ehrlichia chaffeensis and Its Invasin EtpE Block Reactive Oxygen Species Generation by Macrophages in a DNase X-Dependent Manner

    doi: 10.1128/mBio.01551-17

    Figure Lengend Snippet: E. chaffeensis blocks PMA-induced ROS generation by BMDMs from WT mice but not by BMDMs from DNase X −/− mice. BMDMs from WT (A, B) and DNase X −/− (C, D) mice were preincubated with luminol in HBSSd for 15 min and then incubated with E. chaffeensis ( Ech ) isolated from infected DH82 cells, DH82 cell lysate, or HBSSd (control [CTL]) at 37°C for 30 min. ROS generation was continuously recorded as the relative chemiluminescence of oxidized luminol after the addition of PMA (0.5 µg/ml, indicated by arrows) (A, C). UN, unstimulated BMDMs in HBSSd without PMA addition. The area under the curve was measured over 60 min after PMA addition and is shown relative to ROS generation in the control with PMA, which was considered 100% (B, D). Results are presented as the mean ± the standard deviation from at least three independent experiments and were compared with a Student t test; *, P

    Article Snippet: Reactive bands were visualized by enhanced chemiluminescence by incubating the membrane with the respective horseradish peroxidase-conjugated secondary antibodies and the same volume of H2 O2 solution (Thermo Scientific) and Luminol/Enhancer solution (Thermo Scientific).

    Techniques: Mouse Assay, Incubation, Isolation, Infection, CTL Assay, Standard Deviation