goat anti mouse igg texas red  (Thermo Fisher)


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    Goat anti Mouse IgG Texas Red
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    Thermo Fisher goat anti mouse igg texas red
    <t>MBL</t> binds to B. burgdorferi , but MBL deficiency in mice does not decrease complement-dependent killing of B. burgdorferi . ( A ) Recombinant human MBL (5 µg/mL) was added to cultured CFSE-labeled B. burgdorferi N40 spirochetes (2 × 10 8 spirochetes/mL) and after two washing steps, binding of MBL to B. burgdorferi was subjected to FACS analysis (red line). As a control, 100 mM EDTA was added together with MBL which abrogates calcium dependent binding of MBL CRD’s (yellow dotted line). The blue line represents spirochetes without MBL. Binding of MBL was determined using a mouse anti-human MBL antibody and a goat anti-mouse <t>IgG</t> Texas Red antibody. ( B ) ELISA assessment of dose-dependent binding of MBL in murine serum to B. burgdorferi membrane protein extract-coated plates. WT serum or MBL deficient murine serum was added to wells and murine MBL was detected using an antibody against murine MBL-A. As a control, normal murine serum was pre-incubated with 100 mM EDTA. ( C ) Growth inhibition of B. burgdorferi strain N40 by murine serum was measured by the color change of Phenol Red at 562/630 nm once a day for up to five days. Ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 3.66 and 3.01, respectively. ( D ) Growth inhibition of B. garinii strain A87S by murine serum. The ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 2.84 and 2.86, respectively. ( E ) B. garinii strain A87S was incubated with 25% WT NMS, 25% MBL deficient murine serum or 25% heat-inactivated (HI) NMS (control). After 1 h of incubation the percentages of non-motile spirochetes were determined by a researcher blinded to the experimental design. Each sample and each time point were performed in triplicates and symbols or bars represent the mean ± SEM.

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    1) Product Images from "The role of Mannose Binding Lectin in the immune response against Borrelia burgdorferi sensu lato"

    Article Title: The role of Mannose Binding Lectin in the immune response against Borrelia burgdorferi sensu lato

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-37922-8

    MBL binds to B. burgdorferi , but MBL deficiency in mice does not decrease complement-dependent killing of B. burgdorferi . ( A ) Recombinant human MBL (5 µg/mL) was added to cultured CFSE-labeled B. burgdorferi N40 spirochetes (2 × 10 8 spirochetes/mL) and after two washing steps, binding of MBL to B. burgdorferi was subjected to FACS analysis (red line). As a control, 100 mM EDTA was added together with MBL which abrogates calcium dependent binding of MBL CRD’s (yellow dotted line). The blue line represents spirochetes without MBL. Binding of MBL was determined using a mouse anti-human MBL antibody and a goat anti-mouse IgG Texas Red antibody. ( B ) ELISA assessment of dose-dependent binding of MBL in murine serum to B. burgdorferi membrane protein extract-coated plates. WT serum or MBL deficient murine serum was added to wells and murine MBL was detected using an antibody against murine MBL-A. As a control, normal murine serum was pre-incubated with 100 mM EDTA. ( C ) Growth inhibition of B. burgdorferi strain N40 by murine serum was measured by the color change of Phenol Red at 562/630 nm once a day for up to five days. Ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 3.66 and 3.01, respectively. ( D ) Growth inhibition of B. garinii strain A87S by murine serum. The ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 2.84 and 2.86, respectively. ( E ) B. garinii strain A87S was incubated with 25% WT NMS, 25% MBL deficient murine serum or 25% heat-inactivated (HI) NMS (control). After 1 h of incubation the percentages of non-motile spirochetes were determined by a researcher blinded to the experimental design. Each sample and each time point were performed in triplicates and symbols or bars represent the mean ± SEM.
    Figure Legend Snippet: MBL binds to B. burgdorferi , but MBL deficiency in mice does not decrease complement-dependent killing of B. burgdorferi . ( A ) Recombinant human MBL (5 µg/mL) was added to cultured CFSE-labeled B. burgdorferi N40 spirochetes (2 × 10 8 spirochetes/mL) and after two washing steps, binding of MBL to B. burgdorferi was subjected to FACS analysis (red line). As a control, 100 mM EDTA was added together with MBL which abrogates calcium dependent binding of MBL CRD’s (yellow dotted line). The blue line represents spirochetes without MBL. Binding of MBL was determined using a mouse anti-human MBL antibody and a goat anti-mouse IgG Texas Red antibody. ( B ) ELISA assessment of dose-dependent binding of MBL in murine serum to B. burgdorferi membrane protein extract-coated plates. WT serum or MBL deficient murine serum was added to wells and murine MBL was detected using an antibody against murine MBL-A. As a control, normal murine serum was pre-incubated with 100 mM EDTA. ( C ) Growth inhibition of B. burgdorferi strain N40 by murine serum was measured by the color change of Phenol Red at 562/630 nm once a day for up to five days. Ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 3.66 and 3.01, respectively. ( D ) Growth inhibition of B. garinii strain A87S by murine serum. The ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 2.84 and 2.86, respectively. ( E ) B. garinii strain A87S was incubated with 25% WT NMS, 25% MBL deficient murine serum or 25% heat-inactivated (HI) NMS (control). After 1 h of incubation the percentages of non-motile spirochetes were determined by a researcher blinded to the experimental design. Each sample and each time point were performed in triplicates and symbols or bars represent the mean ± SEM.

    Techniques Used: Mouse Assay, Recombinant, Cell Culture, Labeling, Binding Assay, FACS, Enzyme-linked Immunosorbent Assay, Incubation, Inhibition

    Increased B. burgdorferi burden in skin tissue but not in other tissues of MBL deficient mice during murine B. burgdorferi infection. ( A – C ) WT and MBL deficient mice (8 per group) were infected with 10 6 B. burgdorferi N40 via subcutaneous inoculation. Mice were sacrificed after 14 days and assessed for B. burgdorferi burden. DNA was extracted using the Qiagen Blood and Tissue kit from skin, heart, joint and bladder and were subjected to quantitative B. burgdorferi flaB PCR normalized for quantitative mouse β-actin PCR. A. qPCR assessment of B. burgdorferi burden in skin after 14 days. ( B ) IgG and IgM titers against B. burgdorferi lysate in serum measured by ELISA. ( C ) qPCR assessment of B. burgdorferi burden in joint, bladder or heart. ( D,E ) WT and MBL deficient mice (8 per group) were infected with B. burgdorferi N40 via a tick challenge (4–5 ticks/mouse). Two separate experiments were performed, one with three animals per group and one with five animals per group. B. burgdorferi burden was normalized to the highest WT control value per experiment to pool both experiments. ( D ) qPCR assessment of B. burgdorferi burden in skin from ear biopsy at 7 days. ( E ) qPCR assessment of B. burgdorferi burden in skin, joint, bladder or heart at 21 days. ( F,G ) Five µm-thick sagittal heart sections were stained with hematoxylin and eosin. A carditis score was performed by an independent pathologist who was blinded to the experimental design on a scale from 0–3 with 0 being no, 1 mild, 2 moderate, and 3 being severe carditis. Carditis was characterized by disperse inflammation at the atrioventricular junction and aortic root. See Fig. S1 for a normal heart from a mice injected with PBS as a control. Error bars represent mean ± SEM and mean values significantly different in a two-tailed non-parametric Mann-Whitney test are indicated by asterisks (*p
    Figure Legend Snippet: Increased B. burgdorferi burden in skin tissue but not in other tissues of MBL deficient mice during murine B. burgdorferi infection. ( A – C ) WT and MBL deficient mice (8 per group) were infected with 10 6 B. burgdorferi N40 via subcutaneous inoculation. Mice were sacrificed after 14 days and assessed for B. burgdorferi burden. DNA was extracted using the Qiagen Blood and Tissue kit from skin, heart, joint and bladder and were subjected to quantitative B. burgdorferi flaB PCR normalized for quantitative mouse β-actin PCR. A. qPCR assessment of B. burgdorferi burden in skin after 14 days. ( B ) IgG and IgM titers against B. burgdorferi lysate in serum measured by ELISA. ( C ) qPCR assessment of B. burgdorferi burden in joint, bladder or heart. ( D,E ) WT and MBL deficient mice (8 per group) were infected with B. burgdorferi N40 via a tick challenge (4–5 ticks/mouse). Two separate experiments were performed, one with three animals per group and one with five animals per group. B. burgdorferi burden was normalized to the highest WT control value per experiment to pool both experiments. ( D ) qPCR assessment of B. burgdorferi burden in skin from ear biopsy at 7 days. ( E ) qPCR assessment of B. burgdorferi burden in skin, joint, bladder or heart at 21 days. ( F,G ) Five µm-thick sagittal heart sections were stained with hematoxylin and eosin. A carditis score was performed by an independent pathologist who was blinded to the experimental design on a scale from 0–3 with 0 being no, 1 mild, 2 moderate, and 3 being severe carditis. Carditis was characterized by disperse inflammation at the atrioventricular junction and aortic root. See Fig. S1 for a normal heart from a mice injected with PBS as a control. Error bars represent mean ± SEM and mean values significantly different in a two-tailed non-parametric Mann-Whitney test are indicated by asterisks (*p

    Techniques Used: Mouse Assay, Infection, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining, Injection, Two Tailed Test, MANN-WHITNEY

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    Staining:

    Article Title: Expression of Legionella pneumophila Virulence Traits in Response to Growth Conditions
    Article Snippet: Macrophages cultured on coverslips were infected for 1 h, washed twice to remove the majority of extracellular bacteria, and then incubated in fresh medium for various periods of time. .. Duplicate samples were fixed and stained essentially as described previously , using the following reagents: supernatant containing a mouse anti- L. pneumophila flagellum monoclonal antibody (a gift from N. C. Engleberg) was diluted 1:100; Texas red-conjugated goat anti-mouse immunoglobulin G (Molecular Probes) was diluted 1:1,500; and bacteria were stained with 0.1 μg of the DNA stain 4,6-diamidino-2-phenylindole (DAPI; Sigma) per ml of PBS. .. Preparations were examined with a Zeiss Axioplan 2 microscope equipped with a 100× Plan-Neofluar objective lens with a numerical aperture of 1.3.

    Article Title: Filamin A-Hinge Region 1-EGFP: A Novel Tool for Tracking the Cellular Functions of Filamin A in Real-Time
    Article Snippet: Immunostaining and confocal microscopy To analyze the subcellular localization of FLNa-EGFP, A7 and M2 cells were seeded on coverslips and M2 cells were transiently transfected with pcDNA3.1-FLNa-EGFP or pREP4-FLNa using Fugene 6. .. Cells were then washed in PBS, fixed and permeabilized in PBS with 0.1% Tween 20 before staining with mouse anti-filamin 1 (Santa Cruz Biotechnologies, CA, USA), Alexa Fluor 488 goat anti-mouse (Invitrogen, CA, USA) or Texas Red goat anti-mouse (Invitrogen, CA, USA) and Rhodamine-Phalloidin (Invitrogen, CA, USA). .. All coverslips were mounted using Prolong Gold mounting medium with DAPI (Invitrogen, CA, USA).

    Article Title: The Syk inhibitor R406 is a modulator of P-glycoprotein (ABCB1)-mediated multidrug resistance
    Article Snippet: Cells were harvested, exposed to either rhodamine-123 or BODIPY-paclitaxel for 1 h at 37 °C, drug was removed by centrifugation (200 x g ) at 4 °C, and cells washed once in cold PBS. .. In order to correlate drug accumulation with P-gp content, cells were stained on ice using an anti-P-gp mouse monoclonal which detects an external epitope (clone UIC2, EMD Millipore, Billerica, MA), and detected by a Texas Red goat anti-mouse secondary antibody (Thermo Fisher Scientific) using an LSR II flow cytometer. ..

    Article Title: Neuroglobin Regulates Wnt/β-Catenin and NFκB Signaling Pathway through Dvl1
    Article Snippet: .. After 24 h post-transfection, cells were fixed by 4% paraformaldehyde and co-incubated with rabbit polyclonal antibodies against HA-tag and mouse polyclonal antibodies against Myc-tag for 4 h. Green-conjugated anti-rabbit IgG and Red-conjugated anti-mouse IgG were added into cells and incubated for 2 h. Nucleus was stained with Hoechst 33258. ..

    Immunofluorescence:

    Article Title: Deletion of Ezrin in B cells of Lyn-deficient mice downregulates lupus pathology
    Article Snippet: For light microscopy, sections were cut and stained with hematoxylin and eosin (H & E). .. IgG immune complex and complement C3 deposits in kidney were detected by direct immunofluorescence of OCT-embedded frozen kidney sections (5 μm thick) using Texas Red-conjugated goat anti-mouse IgG (Molecular Probes) or FITC-conjugated anti-C3 complement (Thermo Fisher Scientific), respectively. .. The slides were mounted in Vectashield (Vector Laboratories) and observed under the microscope.

    Flow Cytometry:

    Article Title: The Syk inhibitor R406 is a modulator of P-glycoprotein (ABCB1)-mediated multidrug resistance
    Article Snippet: Cells were harvested, exposed to either rhodamine-123 or BODIPY-paclitaxel for 1 h at 37 °C, drug was removed by centrifugation (200 x g ) at 4 °C, and cells washed once in cold PBS. .. In order to correlate drug accumulation with P-gp content, cells were stained on ice using an anti-P-gp mouse monoclonal which detects an external epitope (clone UIC2, EMD Millipore, Billerica, MA), and detected by a Texas Red goat anti-mouse secondary antibody (Thermo Fisher Scientific) using an LSR II flow cytometer. ..

    Cytometry:

    Article Title: The Syk inhibitor R406 is a modulator of P-glycoprotein (ABCB1)-mediated multidrug resistance
    Article Snippet: Cells were harvested, exposed to either rhodamine-123 or BODIPY-paclitaxel for 1 h at 37 °C, drug was removed by centrifugation (200 x g ) at 4 °C, and cells washed once in cold PBS. .. In order to correlate drug accumulation with P-gp content, cells were stained on ice using an anti-P-gp mouse monoclonal which detects an external epitope (clone UIC2, EMD Millipore, Billerica, MA), and detected by a Texas Red goat anti-mouse secondary antibody (Thermo Fisher Scientific) using an LSR II flow cytometer. ..

    Incubation:

    Article Title: Neuroglobin Regulates Wnt/β-Catenin and NFκB Signaling Pathway through Dvl1
    Article Snippet: .. After 24 h post-transfection, cells were fixed by 4% paraformaldehyde and co-incubated with rabbit polyclonal antibodies against HA-tag and mouse polyclonal antibodies against Myc-tag for 4 h. Green-conjugated anti-rabbit IgG and Red-conjugated anti-mouse IgG were added into cells and incubated for 2 h. Nucleus was stained with Hoechst 33258. ..

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    Thermo Fisher goat anti mouse igg texas red
    <t>MBL</t> binds to B. burgdorferi , but MBL deficiency in mice does not decrease complement-dependent killing of B. burgdorferi . ( A ) Recombinant human MBL (5 µg/mL) was added to cultured CFSE-labeled B. burgdorferi N40 spirochetes (2 × 10 8 spirochetes/mL) and after two washing steps, binding of MBL to B. burgdorferi was subjected to FACS analysis (red line). As a control, 100 mM EDTA was added together with MBL which abrogates calcium dependent binding of MBL CRD’s (yellow dotted line). The blue line represents spirochetes without MBL. Binding of MBL was determined using a mouse anti-human MBL antibody and a goat anti-mouse <t>IgG</t> Texas Red antibody. ( B ) ELISA assessment of dose-dependent binding of MBL in murine serum to B. burgdorferi membrane protein extract-coated plates. WT serum or MBL deficient murine serum was added to wells and murine MBL was detected using an antibody against murine MBL-A. As a control, normal murine serum was pre-incubated with 100 mM EDTA. ( C ) Growth inhibition of B. burgdorferi strain N40 by murine serum was measured by the color change of Phenol Red at 562/630 nm once a day for up to five days. Ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 3.66 and 3.01, respectively. ( D ) Growth inhibition of B. garinii strain A87S by murine serum. The ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 2.84 and 2.86, respectively. ( E ) B. garinii strain A87S was incubated with 25% WT NMS, 25% MBL deficient murine serum or 25% heat-inactivated (HI) NMS (control). After 1 h of incubation the percentages of non-motile spirochetes were determined by a researcher blinded to the experimental design. Each sample and each time point were performed in triplicates and symbols or bars represent the mean ± SEM.
    Goat Anti Mouse Igg Texas Red, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher mouse anti α tubulin antibody
    (A) H 2 O 2 -induced accumulation of hOxr1 protein in HeLa cells. Cells were loaded with 100 μM MitoTracker Red for 15 min and then treated with 1 mM H 2 O 2 for 15 min or left untreated and allowed to recover in fresh medium for 1 h prior to fixation and staining with anti-C7C antibody. (B) HeLa cells were treated with 1 mM H 2 O 2 for 15 min and allowed to recover in fresh medium for the times indicated. Cells were harvested, and crude protein extracts were separated by SDS-polyacrylamide gel electrophoresis and immunoblotted with anti-C7C antibody. Samples were also stained with Coomassie as a loading control. (C) Cells were heat stressed at 42°C for 30 min and allowed to recover at 37°C for the times indicated. Western blot assays were conducted as in panel B. Blots were probed with <t>anti-α-tubulin</t> antibody as a loading control. U, unstressed.
    Mouse Anti α Tubulin Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher ed 1 positive macrophages
    Macrophage activation induces axon regeneration in the rat optic nerve. Sections through the retina ( a , c , e ) or the optic nerve ( b , d , f ) were stained with antibodies to detect GAP-43 ( a – f , green fluorescence) or the macrophage marker <t>ED-1</t> ( a , c , e , red fluorescence) 2 weeks after optic nerve surgery. a , GAP-43 is not detected in the GCL ( open arrowheads ) of animals that had received control PBS injections after optic nerve damage; ED-1 + macrophages are absent. b , Few GAP-43-positive axons extend past the injury site ( asterisk ) in the optic nerve. c , Zymosan injected into the vitreous the same day as nerve injury stimulates ED-1 + macrophages to infiltrate the eye and distribute near the GCL ( arrows ). GAP-43 expression is intense in RGC somata ( arrowheads ), and many GAP-43-positive axons extend into the distal optic nerve ( d ). e , Zymosan injections made 3 d after nerve injury result in high levels of GAP-43 in RGCs ( arrowheads ) and greater numbers of GAP-43-positive axons extending distal to the injury site ( f ). Scale bars: a , c , e , 250 μm; b , d , f , 200 μm.
    Ed 1 Positive Macrophages, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MBL binds to B. burgdorferi , but MBL deficiency in mice does not decrease complement-dependent killing of B. burgdorferi . ( A ) Recombinant human MBL (5 µg/mL) was added to cultured CFSE-labeled B. burgdorferi N40 spirochetes (2 × 10 8 spirochetes/mL) and after two washing steps, binding of MBL to B. burgdorferi was subjected to FACS analysis (red line). As a control, 100 mM EDTA was added together with MBL which abrogates calcium dependent binding of MBL CRD’s (yellow dotted line). The blue line represents spirochetes without MBL. Binding of MBL was determined using a mouse anti-human MBL antibody and a goat anti-mouse IgG Texas Red antibody. ( B ) ELISA assessment of dose-dependent binding of MBL in murine serum to B. burgdorferi membrane protein extract-coated plates. WT serum or MBL deficient murine serum was added to wells and murine MBL was detected using an antibody against murine MBL-A. As a control, normal murine serum was pre-incubated with 100 mM EDTA. ( C ) Growth inhibition of B. burgdorferi strain N40 by murine serum was measured by the color change of Phenol Red at 562/630 nm once a day for up to five days. Ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 3.66 and 3.01, respectively. ( D ) Growth inhibition of B. garinii strain A87S by murine serum. The ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 2.84 and 2.86, respectively. ( E ) B. garinii strain A87S was incubated with 25% WT NMS, 25% MBL deficient murine serum or 25% heat-inactivated (HI) NMS (control). After 1 h of incubation the percentages of non-motile spirochetes were determined by a researcher blinded to the experimental design. Each sample and each time point were performed in triplicates and symbols or bars represent the mean ± SEM.

    Journal: Scientific Reports

    Article Title: The role of Mannose Binding Lectin in the immune response against Borrelia burgdorferi sensu lato

    doi: 10.1038/s41598-018-37922-8

    Figure Lengend Snippet: MBL binds to B. burgdorferi , but MBL deficiency in mice does not decrease complement-dependent killing of B. burgdorferi . ( A ) Recombinant human MBL (5 µg/mL) was added to cultured CFSE-labeled B. burgdorferi N40 spirochetes (2 × 10 8 spirochetes/mL) and after two washing steps, binding of MBL to B. burgdorferi was subjected to FACS analysis (red line). As a control, 100 mM EDTA was added together with MBL which abrogates calcium dependent binding of MBL CRD’s (yellow dotted line). The blue line represents spirochetes without MBL. Binding of MBL was determined using a mouse anti-human MBL antibody and a goat anti-mouse IgG Texas Red antibody. ( B ) ELISA assessment of dose-dependent binding of MBL in murine serum to B. burgdorferi membrane protein extract-coated plates. WT serum or MBL deficient murine serum was added to wells and murine MBL was detected using an antibody against murine MBL-A. As a control, normal murine serum was pre-incubated with 100 mM EDTA. ( C ) Growth inhibition of B. burgdorferi strain N40 by murine serum was measured by the color change of Phenol Red at 562/630 nm once a day for up to five days. Ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 3.66 and 3.01, respectively. ( D ) Growth inhibition of B. garinii strain A87S by murine serum. The ratio NMS/HI NMS and MBL deficient serum/HI MBL deficient serum after five days was 2.84 and 2.86, respectively. ( E ) B. garinii strain A87S was incubated with 25% WT NMS, 25% MBL deficient murine serum or 25% heat-inactivated (HI) NMS (control). After 1 h of incubation the percentages of non-motile spirochetes were determined by a researcher blinded to the experimental design. Each sample and each time point were performed in triplicates and symbols or bars represent the mean ± SEM.

    Article Snippet: Spirochetes were washed and bound MBL was detected using mouse anti-human MBL (1:250) (mAb 3E7, Hycult-biotech, Uden, the Netherlands) and goat anti-mouse IgG Texas Red (1:1000, Invitrogen, CA, USA).

    Techniques: Mouse Assay, Recombinant, Cell Culture, Labeling, Binding Assay, FACS, Enzyme-linked Immunosorbent Assay, Incubation, Inhibition

    Increased B. burgdorferi burden in skin tissue but not in other tissues of MBL deficient mice during murine B. burgdorferi infection. ( A – C ) WT and MBL deficient mice (8 per group) were infected with 10 6 B. burgdorferi N40 via subcutaneous inoculation. Mice were sacrificed after 14 days and assessed for B. burgdorferi burden. DNA was extracted using the Qiagen Blood and Tissue kit from skin, heart, joint and bladder and were subjected to quantitative B. burgdorferi flaB PCR normalized for quantitative mouse β-actin PCR. A. qPCR assessment of B. burgdorferi burden in skin after 14 days. ( B ) IgG and IgM titers against B. burgdorferi lysate in serum measured by ELISA. ( C ) qPCR assessment of B. burgdorferi burden in joint, bladder or heart. ( D,E ) WT and MBL deficient mice (8 per group) were infected with B. burgdorferi N40 via a tick challenge (4–5 ticks/mouse). Two separate experiments were performed, one with three animals per group and one with five animals per group. B. burgdorferi burden was normalized to the highest WT control value per experiment to pool both experiments. ( D ) qPCR assessment of B. burgdorferi burden in skin from ear biopsy at 7 days. ( E ) qPCR assessment of B. burgdorferi burden in skin, joint, bladder or heart at 21 days. ( F,G ) Five µm-thick sagittal heart sections were stained with hematoxylin and eosin. A carditis score was performed by an independent pathologist who was blinded to the experimental design on a scale from 0–3 with 0 being no, 1 mild, 2 moderate, and 3 being severe carditis. Carditis was characterized by disperse inflammation at the atrioventricular junction and aortic root. See Fig. S1 for a normal heart from a mice injected with PBS as a control. Error bars represent mean ± SEM and mean values significantly different in a two-tailed non-parametric Mann-Whitney test are indicated by asterisks (*p

    Journal: Scientific Reports

    Article Title: The role of Mannose Binding Lectin in the immune response against Borrelia burgdorferi sensu lato

    doi: 10.1038/s41598-018-37922-8

    Figure Lengend Snippet: Increased B. burgdorferi burden in skin tissue but not in other tissues of MBL deficient mice during murine B. burgdorferi infection. ( A – C ) WT and MBL deficient mice (8 per group) were infected with 10 6 B. burgdorferi N40 via subcutaneous inoculation. Mice were sacrificed after 14 days and assessed for B. burgdorferi burden. DNA was extracted using the Qiagen Blood and Tissue kit from skin, heart, joint and bladder and were subjected to quantitative B. burgdorferi flaB PCR normalized for quantitative mouse β-actin PCR. A. qPCR assessment of B. burgdorferi burden in skin after 14 days. ( B ) IgG and IgM titers against B. burgdorferi lysate in serum measured by ELISA. ( C ) qPCR assessment of B. burgdorferi burden in joint, bladder or heart. ( D,E ) WT and MBL deficient mice (8 per group) were infected with B. burgdorferi N40 via a tick challenge (4–5 ticks/mouse). Two separate experiments were performed, one with three animals per group and one with five animals per group. B. burgdorferi burden was normalized to the highest WT control value per experiment to pool both experiments. ( D ) qPCR assessment of B. burgdorferi burden in skin from ear biopsy at 7 days. ( E ) qPCR assessment of B. burgdorferi burden in skin, joint, bladder or heart at 21 days. ( F,G ) Five µm-thick sagittal heart sections were stained with hematoxylin and eosin. A carditis score was performed by an independent pathologist who was blinded to the experimental design on a scale from 0–3 with 0 being no, 1 mild, 2 moderate, and 3 being severe carditis. Carditis was characterized by disperse inflammation at the atrioventricular junction and aortic root. See Fig. S1 for a normal heart from a mice injected with PBS as a control. Error bars represent mean ± SEM and mean values significantly different in a two-tailed non-parametric Mann-Whitney test are indicated by asterisks (*p

    Article Snippet: Spirochetes were washed and bound MBL was detected using mouse anti-human MBL (1:250) (mAb 3E7, Hycult-biotech, Uden, the Netherlands) and goat anti-mouse IgG Texas Red (1:1000, Invitrogen, CA, USA).

    Techniques: Mouse Assay, Infection, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining, Injection, Two Tailed Test, MANN-WHITNEY

    FLNa-EGFP colocalizes with actin at the membrane and in fibres. ( A ) Confocal images of FLNa, FLNa-EGFP and actin expression in cells after immunostaining. M2 cells transiently transfected with pcDNA3.1-FLNa-EGFP or pREP4-FLNa and untransfected A7 and M2 cells, were seeded on top of glass coverslips, fixed, permeabilized, incubated with anti-FLNa and goat anti-mouse Alexa Fluor 488 and stained with rhodamine-phalloidin before mounting. ( B ) Confocal images of FLNa-EGFP expression in M2 cells transiently transfected with pcDNA3.1-FLNa-EGFP. Cells were seeded on top of glass coverslips, fixed, permeabilized and incubated with anti-FLNa and Texas Red goat anti-mouse. All images are from one single layer of the Z stacks. The colocalization between (green) and (red) was analyzed using Imaris colocalization software and is shown in white. Images are representative of at least three independent experiments. Scalebars (10 μm).

    Journal: PLoS ONE

    Article Title: Filamin A-Hinge Region 1-EGFP: A Novel Tool for Tracking the Cellular Functions of Filamin A in Real-Time

    doi: 10.1371/journal.pone.0040864

    Figure Lengend Snippet: FLNa-EGFP colocalizes with actin at the membrane and in fibres. ( A ) Confocal images of FLNa, FLNa-EGFP and actin expression in cells after immunostaining. M2 cells transiently transfected with pcDNA3.1-FLNa-EGFP or pREP4-FLNa and untransfected A7 and M2 cells, were seeded on top of glass coverslips, fixed, permeabilized, incubated with anti-FLNa and goat anti-mouse Alexa Fluor 488 and stained with rhodamine-phalloidin before mounting. ( B ) Confocal images of FLNa-EGFP expression in M2 cells transiently transfected with pcDNA3.1-FLNa-EGFP. Cells were seeded on top of glass coverslips, fixed, permeabilized and incubated with anti-FLNa and Texas Red goat anti-mouse. All images are from one single layer of the Z stacks. The colocalization between (green) and (red) was analyzed using Imaris colocalization software and is shown in white. Images are representative of at least three independent experiments. Scalebars (10 μm).

    Article Snippet: Cells were then washed in PBS, fixed and permeabilized in PBS with 0.1% Tween 20 before staining with mouse anti-filamin 1 (Santa Cruz Biotechnologies, CA, USA), Alexa Fluor 488 goat anti-mouse (Invitrogen, CA, USA) or Texas Red goat anti-mouse (Invitrogen, CA, USA) and Rhodamine-Phalloidin (Invitrogen, CA, USA).

    Techniques: Expressing, Immunostaining, Transfection, Incubation, Staining, Software

    (A) H 2 O 2 -induced accumulation of hOxr1 protein in HeLa cells. Cells were loaded with 100 μM MitoTracker Red for 15 min and then treated with 1 mM H 2 O 2 for 15 min or left untreated and allowed to recover in fresh medium for 1 h prior to fixation and staining with anti-C7C antibody. (B) HeLa cells were treated with 1 mM H 2 O 2 for 15 min and allowed to recover in fresh medium for the times indicated. Cells were harvested, and crude protein extracts were separated by SDS-polyacrylamide gel electrophoresis and immunoblotted with anti-C7C antibody. Samples were also stained with Coomassie as a loading control. (C) Cells were heat stressed at 42°C for 30 min and allowed to recover at 37°C for the times indicated. Western blot assays were conducted as in panel B. Blots were probed with anti-α-tubulin antibody as a loading control. U, unstressed.

    Journal: Molecular and Cellular Biology

    Article Title: Stress Induction and Mitochondrial Localization of Oxr1 Proteins in Yeast and Humans

    doi: 10.1128/MCB.24.8.3180-3187.2004

    Figure Lengend Snippet: (A) H 2 O 2 -induced accumulation of hOxr1 protein in HeLa cells. Cells were loaded with 100 μM MitoTracker Red for 15 min and then treated with 1 mM H 2 O 2 for 15 min or left untreated and allowed to recover in fresh medium for 1 h prior to fixation and staining with anti-C7C antibody. (B) HeLa cells were treated with 1 mM H 2 O 2 for 15 min and allowed to recover in fresh medium for the times indicated. Cells were harvested, and crude protein extracts were separated by SDS-polyacrylamide gel electrophoresis and immunoblotted with anti-C7C antibody. Samples were also stained with Coomassie as a loading control. (C) Cells were heat stressed at 42°C for 30 min and allowed to recover at 37°C for the times indicated. Western blot assays were conducted as in panel B. Blots were probed with anti-α-tubulin antibody as a loading control. U, unstressed.

    Article Snippet: Where shown, mouse anti-α-tubulin antibody (Lab Vision) was used to probe blots as a loading control.

    Techniques: Staining, Polyacrylamide Gel Electrophoresis, Western Blot

    Macrophage activation induces axon regeneration in the rat optic nerve. Sections through the retina ( a , c , e ) or the optic nerve ( b , d , f ) were stained with antibodies to detect GAP-43 ( a – f , green fluorescence) or the macrophage marker ED-1 ( a , c , e , red fluorescence) 2 weeks after optic nerve surgery. a , GAP-43 is not detected in the GCL ( open arrowheads ) of animals that had received control PBS injections after optic nerve damage; ED-1 + macrophages are absent. b , Few GAP-43-positive axons extend past the injury site ( asterisk ) in the optic nerve. c , Zymosan injected into the vitreous the same day as nerve injury stimulates ED-1 + macrophages to infiltrate the eye and distribute near the GCL ( arrows ). GAP-43 expression is intense in RGC somata ( arrowheads ), and many GAP-43-positive axons extend into the distal optic nerve ( d ). e , Zymosan injections made 3 d after nerve injury result in high levels of GAP-43 in RGCs ( arrowheads ) and greater numbers of GAP-43-positive axons extending distal to the injury site ( f ). Scale bars: a , c , e , 250 μm; b , d , f , 200 μm.

    Journal: The Journal of Neuroscience

    Article Title: Macrophage-Derived Factors Stimulate Optic Nerve Regeneration

    doi: 10.1523/JNEUROSCI.23-06-02284.2003

    Figure Lengend Snippet: Macrophage activation induces axon regeneration in the rat optic nerve. Sections through the retina ( a , c , e ) or the optic nerve ( b , d , f ) were stained with antibodies to detect GAP-43 ( a – f , green fluorescence) or the macrophage marker ED-1 ( a , c , e , red fluorescence) 2 weeks after optic nerve surgery. a , GAP-43 is not detected in the GCL ( open arrowheads ) of animals that had received control PBS injections after optic nerve damage; ED-1 + macrophages are absent. b , Few GAP-43-positive axons extend past the injury site ( asterisk ) in the optic nerve. c , Zymosan injected into the vitreous the same day as nerve injury stimulates ED-1 + macrophages to infiltrate the eye and distribute near the GCL ( arrows ). GAP-43 expression is intense in RGC somata ( arrowheads ), and many GAP-43-positive axons extend into the distal optic nerve ( d ). e , Zymosan injections made 3 d after nerve injury result in high levels of GAP-43 in RGCs ( arrowheads ) and greater numbers of GAP-43-positive axons extending distal to the injury site ( f ). Scale bars: a , c , e , 250 μm; b , d , f , 200 μm.

    Article Snippet: Secondary antibodies conjugated to distinct fluorophores were used to visualize ED-1-positive macrophages (Texas Red-conjugated anti-mouse IgG made in goat, 1:500; Molecular Probes) and GAP-43-positive RGCs (Alexa Fluor 488-conjugated anti-sheep IgG made in donkey, 1:500; Molecular Probes) in the same sections.

    Techniques: Activation Assay, Staining, Fluorescence, Marker, Injection, Expressing

    Effect of Zymosan dosage on RGC survival. a , Zymosan injections, either on the day of nerve crush ( D0 ) or 3 d later ( D3 ), increased the number of TUJ1 + cells in the retina 3 weeks after a peripheral nerve graft. Zymosan resulted in more surviving TUJ1 + cells when injected at a low concentration (1.25 μg/μl) than at a high concentration (12.5 μg/μl). b , The higher dosage of Zymosan diminished retinal size. c , Normal retina (flat mounted). No macrophages appear in the vitreous or around the GCL, although some ED-1 + microglia are seen ( arrowhead ). d , Axotomy followed by a PN graft increases the number of ED-1 + monocytes ( arrow, arrowheads ) in the retina only slightly. e , Zymosan injections (1.25 μg/μl) result in accumulation of ED-1 + macrophages ( arrows ) in the vitreous and around the GCL. * p

    Journal: The Journal of Neuroscience

    Article Title: Macrophage-Derived Factors Stimulate Optic Nerve Regeneration

    doi: 10.1523/JNEUROSCI.23-06-02284.2003

    Figure Lengend Snippet: Effect of Zymosan dosage on RGC survival. a , Zymosan injections, either on the day of nerve crush ( D0 ) or 3 d later ( D3 ), increased the number of TUJ1 + cells in the retina 3 weeks after a peripheral nerve graft. Zymosan resulted in more surviving TUJ1 + cells when injected at a low concentration (1.25 μg/μl) than at a high concentration (12.5 μg/μl). b , The higher dosage of Zymosan diminished retinal size. c , Normal retina (flat mounted). No macrophages appear in the vitreous or around the GCL, although some ED-1 + microglia are seen ( arrowhead ). d , Axotomy followed by a PN graft increases the number of ED-1 + monocytes ( arrow, arrowheads ) in the retina only slightly. e , Zymosan injections (1.25 μg/μl) result in accumulation of ED-1 + macrophages ( arrows ) in the vitreous and around the GCL. * p

    Article Snippet: Secondary antibodies conjugated to distinct fluorophores were used to visualize ED-1-positive macrophages (Texas Red-conjugated anti-mouse IgG made in goat, 1:500; Molecular Probes) and GAP-43-positive RGCs (Alexa Fluor 488-conjugated anti-sheep IgG made in donkey, 1:500; Molecular Probes) in the same sections.

    Techniques: Injection, Concentration Assay