antibody alexa fluor 594 labeled goat anti rabbit igg antibody  (Thermo Fisher)


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    Name:
    Alexa Fluor 594 Goat Anti Rabbit SFX Kit
    Description:
    Alexa Fluor SFX Kits contain Image iT FX signal enhancer Cat no I36933 plus one of sixteen different Alexa Fluor dye labeled secondary antibodies These kits provide 400 µg 0 2 mL of 2 mg mL of either goat anti mouse IgG or goat anti rabbit IgG antibody as a standard or highly cross adsorbed preparation conjugated to Alexa Fluor 488 Alexa Fluor 555 Alexa Fluor 594 or Alexa Fluor 647 dye four of our most commonly used Alexa Fluor dyes The Alexa Fluor 594 goat anti rabbit IgG highly cross adsorbed included with this kit is also available in a 1000 µg unit size Cat no A11037
    Catalog Number:
    a31632
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Antibodies and Secondary Detection|Cell Analysis
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    Structured Review

    Thermo Fisher antibody alexa fluor 594 labeled goat anti rabbit igg antibody
    Intracellular localization of TagB, TagC, and Sha determined by confocal microscopy. ( A ) Z-stack imaging showing the localization of TagB, TagC, and Sha and their respective serine active site mutant variants during interaction with 5637 bladder epithelial cells after 5 h of incubation. SPATEs were detected by <t>Alexa</t> Fluor 594 (white arrowheads, red fluorescence) using anti-mouse secondary antibody and actin was stained by Alexa Fluor 488- phalloidin (green fluorescence). Images are displayed in a 3D section view with large Z-sections in the X-Y direction (R), Z-projection in the X–Z direction (P), and Z-projection in the Y–Z direction (Q). The green and red lines in R indicate the orthogonal planes of the X–Z and Y–Z projection. For each selected section, the signal was gathered from a span of 5 μm. Scale bar: 10 µm ( B ) Quantitative analysis of fluorescence intensity of F-actin in the cells treated with native or mutant SPATEs. Analysis of fluorescence intensity was done in green channel by measuring the mean gray value on ImageJ. Data represent the mean ± SEM of at least three independent experiments. Significant differences between fluorescence intensity of each native and mutant SPATE treated cell was determined using Student’s t-test with ** p
    Alexa Fluor SFX Kits contain Image iT FX signal enhancer Cat no I36933 plus one of sixteen different Alexa Fluor dye labeled secondary antibodies These kits provide 400 µg 0 2 mL of 2 mg mL of either goat anti mouse IgG or goat anti rabbit IgG antibody as a standard or highly cross adsorbed preparation conjugated to Alexa Fluor 488 Alexa Fluor 555 Alexa Fluor 594 or Alexa Fluor 647 dye four of our most commonly used Alexa Fluor dyes The Alexa Fluor 594 goat anti rabbit IgG highly cross adsorbed included with this kit is also available in a 1000 µg unit size Cat no A11037
    https://www.bioz.com/result/antibody alexa fluor 594 labeled goat anti rabbit igg antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibody alexa fluor 594 labeled goat anti rabbit igg antibody - by Bioz Stars, 2021-03
    97/100 stars

    Images

    1) Product Images from "The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption"

    Article Title: The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21093047

    Intracellular localization of TagB, TagC, and Sha determined by confocal microscopy. ( A ) Z-stack imaging showing the localization of TagB, TagC, and Sha and their respective serine active site mutant variants during interaction with 5637 bladder epithelial cells after 5 h of incubation. SPATEs were detected by Alexa Fluor 594 (white arrowheads, red fluorescence) using anti-mouse secondary antibody and actin was stained by Alexa Fluor 488- phalloidin (green fluorescence). Images are displayed in a 3D section view with large Z-sections in the X-Y direction (R), Z-projection in the X–Z direction (P), and Z-projection in the Y–Z direction (Q). The green and red lines in R indicate the orthogonal planes of the X–Z and Y–Z projection. For each selected section, the signal was gathered from a span of 5 μm. Scale bar: 10 µm ( B ) Quantitative analysis of fluorescence intensity of F-actin in the cells treated with native or mutant SPATEs. Analysis of fluorescence intensity was done in green channel by measuring the mean gray value on ImageJ. Data represent the mean ± SEM of at least three independent experiments. Significant differences between fluorescence intensity of each native and mutant SPATE treated cell was determined using Student’s t-test with ** p
    Figure Legend Snippet: Intracellular localization of TagB, TagC, and Sha determined by confocal microscopy. ( A ) Z-stack imaging showing the localization of TagB, TagC, and Sha and their respective serine active site mutant variants during interaction with 5637 bladder epithelial cells after 5 h of incubation. SPATEs were detected by Alexa Fluor 594 (white arrowheads, red fluorescence) using anti-mouse secondary antibody and actin was stained by Alexa Fluor 488- phalloidin (green fluorescence). Images are displayed in a 3D section view with large Z-sections in the X-Y direction (R), Z-projection in the X–Z direction (P), and Z-projection in the Y–Z direction (Q). The green and red lines in R indicate the orthogonal planes of the X–Z and Y–Z projection. For each selected section, the signal was gathered from a span of 5 μm. Scale bar: 10 µm ( B ) Quantitative analysis of fluorescence intensity of F-actin in the cells treated with native or mutant SPATEs. Analysis of fluorescence intensity was done in green channel by measuring the mean gray value on ImageJ. Data represent the mean ± SEM of at least three independent experiments. Significant differences between fluorescence intensity of each native and mutant SPATE treated cell was determined using Student’s t-test with ** p

    Techniques Used: Confocal Microscopy, Imaging, Mutagenesis, Incubation, Fluorescence, Staining

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    Incubation:

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    Article Snippet: Lowering the ion beam current and electron beam voltage FIB-SEM, slice and view was performed to cyclically perform FIB milling and SEM imaging to image the sample and locate relevant sites with visible nanowires. γ-H2AX Labelling : Cells were cultured on nanowire and control substrates placed in 24 well plates for 96 h. Cells were fixed for 20 min in 3.7% paraformaldehyde, washed with PBS and labelled for 90 min with rabbit anti- γ-H2AX (Cell Signalling Tech, Danvers, USA) (1:200) in PBS with 1% (v/v) Tween 20 and 1% (w/v) BSA (Sigma-Aldrich, St. Louis, USA) at room temperature. .. The samples were washed with PBS and incubated for 90 min with Alexa Fluor 594-goat-anti-rabbit (Invitrogen, Carlsbad, USA) (10 μg/mL) and FITC-labelled phalloidin (Sigma-Aldrich, St. Louis, USA) (1.7 μg/μL) in PBS with 1% (v/v) Tween-20 and 1% (w/v) BSA at room temperature. .. Finally the samples were washed with PBS and stained for 1 min using bisbenzimide (Hoechst 33342, Invitrogen, Carlsbad, USA) at a concentration of 1 μg/mL in PBS.

    other:

    Article Title: ZNF265--a novel spliceosomal protein able to induce alternative splicing
    Article Snippet: Secondary antibodies used were: Alexa Fluor 488–conjugated goat anti–mouse IgG (Molecular Probes), Alexa Fluor 594–conjugated goat anti–rabbit IgG (Molecular Probes), alkaline phosphatase–conjugated rabbit anti–mouse IgG (Sigma-Aldrich), and alkaline phosphatase–conjugated goat anti–rabbit IgG (Sigma-Aldrich).

    Microscopy:

    Article Title: Ligand independent activation of c-Met by fibronectin and ?5?1-integrin regulates ovarian cancer invasion and metastasis
    Article Snippet: .. The samples were washed and incubated with Alexa Fluor 594-labelled goat anti rabbit antibody (1:1000) and fluorescein-labelled goat anti mouse antibody (1:1000), mounted with Prolong Gold (Invitrogen, Carlsbad, CA) and analyzed using a Leica SP2 laser scanning confocal microscope. .. Transient transfection HeyA8 cells were transfected with 30nM siRNA against α5 -integrin (Ambion) or against c-Met using Lipofectamine 2000 transfection reagent (Invitrogen).

    Avidin-Biotin Assay:

    Article Title: Isolation, Characterization, and Transplantation of Cardiac Endothelial Cells
    Article Snippet: The following primary antibody were used: fluorescein isothiocyanate (FITC) antimouse CD31 (1 : 50, clone 390, BD Biosciences, San Jose, CA or eBioscience, San Diego, CA), biotin antimouse Sca-1 (1 : 500, clone D7, BD Biosciences or 1 : 400, eBioscience), purified rat anti mouse CD34 (1 : 50, clone RAM34, BD Biosciences), polyclonal rabbit anti human vWF (1 : 100, Dako, Carpinteria, CA), rabbit polyclonal Ab to eNOS (1 : 100, Abcam, Cambridge, MA), and rabbit polyclonal Ab to Caveolin 1 (1 : 300, Abcam), monoclonal anti-α smooth muscle actin Cy3 (1 : 400, clone 1A4, Sigma, St. Louis, MO), rabbit polyclonal Anti-NG2 Chondroitin Sulfate Proteoglycan (1 : 200, Chemicon, Billerica, MA), rabbit polyclonal anti-GFP (1 : 100, Abcam, Cambridge, MA). .. The following secondary antibodies were used: Avidin-Texas red (1 : 500, Vector), Alexa Fluor 594 chicken antirat IgG (1 : 1000, Invitrogen, Carlbad, CA), Streptavidin-Alexa fluor 594 conjugate (1 : 400, Invitrogen), Alexa 647 goat anti-rabbit IgG (1 : 1000, Invitrogen), Alexa 488 goat anti-rabbit IgG (1 : 1000, Invitrogen), and Alexa 594 goat anti-rabbit IgG (1 : 1000, Invitrogen). .. Tissues and cells were also stained with 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI) to visualize the nuclei and examined by Axiovert 200 fluorescence microscopy (Zeiss, Thornwood, NY).

    Plasmid Preparation:

    Article Title: Isolation, Characterization, and Transplantation of Cardiac Endothelial Cells
    Article Snippet: The following primary antibody were used: fluorescein isothiocyanate (FITC) antimouse CD31 (1 : 50, clone 390, BD Biosciences, San Jose, CA or eBioscience, San Diego, CA), biotin antimouse Sca-1 (1 : 500, clone D7, BD Biosciences or 1 : 400, eBioscience), purified rat anti mouse CD34 (1 : 50, clone RAM34, BD Biosciences), polyclonal rabbit anti human vWF (1 : 100, Dako, Carpinteria, CA), rabbit polyclonal Ab to eNOS (1 : 100, Abcam, Cambridge, MA), and rabbit polyclonal Ab to Caveolin 1 (1 : 300, Abcam), monoclonal anti-α smooth muscle actin Cy3 (1 : 400, clone 1A4, Sigma, St. Louis, MO), rabbit polyclonal Anti-NG2 Chondroitin Sulfate Proteoglycan (1 : 200, Chemicon, Billerica, MA), rabbit polyclonal anti-GFP (1 : 100, Abcam, Cambridge, MA). .. The following secondary antibodies were used: Avidin-Texas red (1 : 500, Vector), Alexa Fluor 594 chicken antirat IgG (1 : 1000, Invitrogen, Carlbad, CA), Streptavidin-Alexa fluor 594 conjugate (1 : 400, Invitrogen), Alexa 647 goat anti-rabbit IgG (1 : 1000, Invitrogen), Alexa 488 goat anti-rabbit IgG (1 : 1000, Invitrogen), and Alexa 594 goat anti-rabbit IgG (1 : 1000, Invitrogen). .. Tissues and cells were also stained with 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI) to visualize the nuclei and examined by Axiovert 200 fluorescence microscopy (Zeiss, Thornwood, NY).

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  • 97
    Thermo Fisher alexa fluor 594 conjugated goat anti rabbit igg
    Localization of TL and GSLII binding sites in T . cruzi . Endocytosis kinetics of fluorescent <t>Alexa</t> <t>Fluor</t> 594 conjugated Tf was performed in order to follow T . cruzi endocytic pathway from the flagellar pocket/cytostome to the reservosomes. Parasites were fixed at different time points and probed with biotinylated TL (A), biotinylated ricin (B) or Alexa 488 conjugated GSLII (C). The addition of chitin hydrolysate clearly shows inhibition of TL and GSLII staining. (A) Co-localization of biotinylated-TL (green) and Tf (red). (B) Co-localization of biotinylated-ricin (green) and Tf (red). Addition of 200 mM galactose abolished the ricin staining. (C) Co-localization of Alexa 488 conjugated GSLII (green) and Tf (red). (D) Co-localization of Alexa 488 conjugated GSLII (green) and TcJ6 (red). (E) Co-localization of Alexa 488 conjugated GSLII (green) and anti-BiP (red). (F) GSLII blotting of cell extracts enriched by GSLII chromatography. GSLII blots of T . cruzi CHAPS- and Triton-soluble (CHAPS+Triton X-114) cell lysate fractions were enriched by GSLII chromatography and then treated (+) or not (-) with PNGase F. Blots were probed with biotinylated-GSLII. The GSLII blot indicates the presence of N- acetylglucosamine modification in both soluble and membrane fractions. Treatment of the fractions with PNGase F decreased the reactivity of GSLII confirming N -glycoprotein type modification.
    Alexa Fluor 594 Conjugated Goat Anti Rabbit Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa fluor 594 conjugated goat anti rabbit igg/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    99
    Thermo Fisher 647 anti rabbit
    Characterization of VH411-S-Tag peptide conjugate binding to the CHO-hLDLR-EGFP cell line. (A) Representative confocal photomicrographs of CHO-hLDLR-EGFP and CHO-hTfR-EGFP cells (green) incubated 1 hr at 37°C with 10 μM of VH411-S-Tag detected post-fixation with an anti-S-Tag and A647-conjugated secondary antibody (cyan) and 10 μg/mL of DiI-LDL. Cell nuclei are labeled with Hoechst#33258 (blue) at 0.5 μg/mL. Co-labeling appears in blue-green/orange in the merged pictures. Note that VH411-S-Tag co-localizes with hLDLR and is internalized by CHO-hLDLR-EGFP cells. (B) Representative confocal photomicrographs of CHO-hLDLR-EGFP cells (green) incubated 1 hr at 37°C with a macromolecular complex (see scheme Fig 3B) resulting from the co-incubation and interaction of 10 μM VH411-S-Tag peptide or VH411Sc-S-Tag peptide with the anti-S-Tag antibody (1/200) and the Alexafluor <t>647-conjugated</t> secondary antibody (1/800) (cyan). Cells were concomitantly exposed to 10 μg/mL of DiI-LDL (red). Cell nuclei are labeled with Hoechst#33258 (blue). Co-labeling appears in blue-green/orange in the merged pictures.
    647 Anti Rabbit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/647 anti rabbit/product/Thermo Fisher
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    97
    Thermo Fisher goat anti rabbit igg alexa fluor 594 conjugated
    Inhibition of the nuclear translocation of NF-κB upon GANT-61 treatment Immunofluorescence analysis was performed on MCF-7 and SK-BR-3 breast cancer cells after treatment with GANT-61 (10 μM) or vehicle alone for 24 hours. After the treatment, the cells were fixed, incubated with an anti-NF-κB antibody, washed, and labeled with an <t>Alexa</t> Fluor-488-conjugated goat anti-mouse <t>IgG</t> antibody. The nuclei were counterstained with Hoechst. Original magnification, 400x.
    Goat Anti Rabbit Igg Alexa Fluor 594 Conjugated, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg alexa fluor 594 conjugated/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg alexa fluor 594 conjugated - by Bioz Stars, 2021-03
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    Image Search Results


    Localization of TL and GSLII binding sites in T . cruzi . Endocytosis kinetics of fluorescent Alexa Fluor 594 conjugated Tf was performed in order to follow T . cruzi endocytic pathway from the flagellar pocket/cytostome to the reservosomes. Parasites were fixed at different time points and probed with biotinylated TL (A), biotinylated ricin (B) or Alexa 488 conjugated GSLII (C). The addition of chitin hydrolysate clearly shows inhibition of TL and GSLII staining. (A) Co-localization of biotinylated-TL (green) and Tf (red). (B) Co-localization of biotinylated-ricin (green) and Tf (red). Addition of 200 mM galactose abolished the ricin staining. (C) Co-localization of Alexa 488 conjugated GSLII (green) and Tf (red). (D) Co-localization of Alexa 488 conjugated GSLII (green) and TcJ6 (red). (E) Co-localization of Alexa 488 conjugated GSLII (green) and anti-BiP (red). (F) GSLII blotting of cell extracts enriched by GSLII chromatography. GSLII blots of T . cruzi CHAPS- and Triton-soluble (CHAPS+Triton X-114) cell lysate fractions were enriched by GSLII chromatography and then treated (+) or not (-) with PNGase F. Blots were probed with biotinylated-GSLII. The GSLII blot indicates the presence of N- acetylglucosamine modification in both soluble and membrane fractions. Treatment of the fractions with PNGase F decreased the reactivity of GSLII confirming N -glycoprotein type modification.

    Journal: PLoS ONE

    Article Title: Specific Endocytosis Blockade of Trypanosoma cruzi Exposed to a Poly-LAcNAc Binding Lectin Suggests that Lectin-Sugar Interactions Participate to Receptor-Mediated Endocytosis

    doi: 10.1371/journal.pone.0163302

    Figure Lengend Snippet: Localization of TL and GSLII binding sites in T . cruzi . Endocytosis kinetics of fluorescent Alexa Fluor 594 conjugated Tf was performed in order to follow T . cruzi endocytic pathway from the flagellar pocket/cytostome to the reservosomes. Parasites were fixed at different time points and probed with biotinylated TL (A), biotinylated ricin (B) or Alexa 488 conjugated GSLII (C). The addition of chitin hydrolysate clearly shows inhibition of TL and GSLII staining. (A) Co-localization of biotinylated-TL (green) and Tf (red). (B) Co-localization of biotinylated-ricin (green) and Tf (red). Addition of 200 mM galactose abolished the ricin staining. (C) Co-localization of Alexa 488 conjugated GSLII (green) and Tf (red). (D) Co-localization of Alexa 488 conjugated GSLII (green) and TcJ6 (red). (E) Co-localization of Alexa 488 conjugated GSLII (green) and anti-BiP (red). (F) GSLII blotting of cell extracts enriched by GSLII chromatography. GSLII blots of T . cruzi CHAPS- and Triton-soluble (CHAPS+Triton X-114) cell lysate fractions were enriched by GSLII chromatography and then treated (+) or not (-) with PNGase F. Blots were probed with biotinylated-GSLII. The GSLII blot indicates the presence of N- acetylglucosamine modification in both soluble and membrane fractions. Treatment of the fractions with PNGase F decreased the reactivity of GSLII confirming N -glycoprotein type modification.

    Article Snippet: Primary antibodies were detected with an Alexa Fluor 594-conjugated goat anti-rabbit IgG (Life Technologies) washed three times in TBS.

    Techniques: Binding Assay, Inhibition, Staining, Chromatography, Modification

    Inhibition of uptake of Tf by TL in epimastigote forms of T . cruzi . Trypanosomes preincubated with biotinylated TL in the presence of 20 μM FMK-024 (25 μg/ml) and in the absence (A, left panel) or presence of competing chitin hydrolysate (A, right panel), were then incubated with Tf Alexa-594 for 5 or 30 min at 27°C. Cells were then fixed and treated for fluorescence microscopy. Similar incubations wherein TL was substituted by GSLII (B) were performed to assess the specificity of the TL labeling. Furthermore, live parasites preincubated with DyLight 488-TL and 20 μM protease inhibitor (FMK-024) for 5 min and then incubated for 60 min in the presence of Alexa Fluor 594 conjugated Tf showed a lectin labeling in the cytostome/cytopharynx (arrowhead), while no Tf labeling (red signal) was observed in these conditions (C, upper panel). In presence of a molar excess of chitin hydrolysate an intense labeling of Tf exclusively concentrate into reservosomes (arrow) while no green signal corresponding to TL was observed anymore (C, lower panel). Inhibition of trypanosomes Tf uptake with TL was furthermore quantified by flow cytometry (D). The TL signal was dropping from 913 to 273 of mfi in the absence or presence of chitin hydrolysate, respectively (D, left histogram). Conversely, Tf signal was increasing from 597 to 3793 of mfi in the absence or presence of chitin hydrolysate, respectively (D, right histogram).

    Journal: PLoS ONE

    Article Title: Specific Endocytosis Blockade of Trypanosoma cruzi Exposed to a Poly-LAcNAc Binding Lectin Suggests that Lectin-Sugar Interactions Participate to Receptor-Mediated Endocytosis

    doi: 10.1371/journal.pone.0163302

    Figure Lengend Snippet: Inhibition of uptake of Tf by TL in epimastigote forms of T . cruzi . Trypanosomes preincubated with biotinylated TL in the presence of 20 μM FMK-024 (25 μg/ml) and in the absence (A, left panel) or presence of competing chitin hydrolysate (A, right panel), were then incubated with Tf Alexa-594 for 5 or 30 min at 27°C. Cells were then fixed and treated for fluorescence microscopy. Similar incubations wherein TL was substituted by GSLII (B) were performed to assess the specificity of the TL labeling. Furthermore, live parasites preincubated with DyLight 488-TL and 20 μM protease inhibitor (FMK-024) for 5 min and then incubated for 60 min in the presence of Alexa Fluor 594 conjugated Tf showed a lectin labeling in the cytostome/cytopharynx (arrowhead), while no Tf labeling (red signal) was observed in these conditions (C, upper panel). In presence of a molar excess of chitin hydrolysate an intense labeling of Tf exclusively concentrate into reservosomes (arrow) while no green signal corresponding to TL was observed anymore (C, lower panel). Inhibition of trypanosomes Tf uptake with TL was furthermore quantified by flow cytometry (D). The TL signal was dropping from 913 to 273 of mfi in the absence or presence of chitin hydrolysate, respectively (D, left histogram). Conversely, Tf signal was increasing from 597 to 3793 of mfi in the absence or presence of chitin hydrolysate, respectively (D, right histogram).

    Article Snippet: Primary antibodies were detected with an Alexa Fluor 594-conjugated goat anti-rabbit IgG (Life Technologies) washed three times in TBS.

    Techniques: Inhibition, Incubation, Fluorescence, Microscopy, Labeling, Protease Inhibitor, Flow Cytometry, Cytometry

    Intracellular localization of TagB, TagC, and Sha determined by confocal microscopy. ( A ) Z-stack imaging showing the localization of TagB, TagC, and Sha and their respective serine active site mutant variants during interaction with 5637 bladder epithelial cells after 5 h of incubation. SPATEs were detected by Alexa Fluor 594 (white arrowheads, red fluorescence) using anti-mouse secondary antibody and actin was stained by Alexa Fluor 488- phalloidin (green fluorescence). Images are displayed in a 3D section view with large Z-sections in the X-Y direction (R), Z-projection in the X–Z direction (P), and Z-projection in the Y–Z direction (Q). The green and red lines in R indicate the orthogonal planes of the X–Z and Y–Z projection. For each selected section, the signal was gathered from a span of 5 μm. Scale bar: 10 µm ( B ) Quantitative analysis of fluorescence intensity of F-actin in the cells treated with native or mutant SPATEs. Analysis of fluorescence intensity was done in green channel by measuring the mean gray value on ImageJ. Data represent the mean ± SEM of at least three independent experiments. Significant differences between fluorescence intensity of each native and mutant SPATE treated cell was determined using Student’s t-test with ** p

    Journal: International Journal of Molecular Sciences

    Article Title: The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

    doi: 10.3390/ijms21093047

    Figure Lengend Snippet: Intracellular localization of TagB, TagC, and Sha determined by confocal microscopy. ( A ) Z-stack imaging showing the localization of TagB, TagC, and Sha and their respective serine active site mutant variants during interaction with 5637 bladder epithelial cells after 5 h of incubation. SPATEs were detected by Alexa Fluor 594 (white arrowheads, red fluorescence) using anti-mouse secondary antibody and actin was stained by Alexa Fluor 488- phalloidin (green fluorescence). Images are displayed in a 3D section view with large Z-sections in the X-Y direction (R), Z-projection in the X–Z direction (P), and Z-projection in the Y–Z direction (Q). The green and red lines in R indicate the orthogonal planes of the X–Z and Y–Z projection. For each selected section, the signal was gathered from a span of 5 μm. Scale bar: 10 µm ( B ) Quantitative analysis of fluorescence intensity of F-actin in the cells treated with native or mutant SPATEs. Analysis of fluorescence intensity was done in green channel by measuring the mean gray value on ImageJ. Data represent the mean ± SEM of at least three independent experiments. Significant differences between fluorescence intensity of each native and mutant SPATE treated cell was determined using Student’s t-test with ** p

    Article Snippet: This was followed by incubation with secondary antibody Alexa Fluor 594-labeled goat anti-rabbit IgG antibody (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Confocal Microscopy, Imaging, Mutagenesis, Incubation, Fluorescence, Staining

    Characterization of VH411-S-Tag peptide conjugate binding to the CHO-hLDLR-EGFP cell line. (A) Representative confocal photomicrographs of CHO-hLDLR-EGFP and CHO-hTfR-EGFP cells (green) incubated 1 hr at 37°C with 10 μM of VH411-S-Tag detected post-fixation with an anti-S-Tag and A647-conjugated secondary antibody (cyan) and 10 μg/mL of DiI-LDL. Cell nuclei are labeled with Hoechst#33258 (blue) at 0.5 μg/mL. Co-labeling appears in blue-green/orange in the merged pictures. Note that VH411-S-Tag co-localizes with hLDLR and is internalized by CHO-hLDLR-EGFP cells. (B) Representative confocal photomicrographs of CHO-hLDLR-EGFP cells (green) incubated 1 hr at 37°C with a macromolecular complex (see scheme Fig 3B) resulting from the co-incubation and interaction of 10 μM VH411-S-Tag peptide or VH411Sc-S-Tag peptide with the anti-S-Tag antibody (1/200) and the Alexafluor 647-conjugated secondary antibody (1/800) (cyan). Cells were concomitantly exposed to 10 μg/mL of DiI-LDL (red). Cell nuclei are labeled with Hoechst#33258 (blue). Co-labeling appears in blue-green/orange in the merged pictures.

    Journal: PLoS ONE

    Article Title: Identification and characterization of highly versatile peptide-vectors that bind non-competitively to the low-density lipoprotein receptor for in vivo targeting and delivery of small molecules and protein cargos

    doi: 10.1371/journal.pone.0191052

    Figure Lengend Snippet: Characterization of VH411-S-Tag peptide conjugate binding to the CHO-hLDLR-EGFP cell line. (A) Representative confocal photomicrographs of CHO-hLDLR-EGFP and CHO-hTfR-EGFP cells (green) incubated 1 hr at 37°C with 10 μM of VH411-S-Tag detected post-fixation with an anti-S-Tag and A647-conjugated secondary antibody (cyan) and 10 μg/mL of DiI-LDL. Cell nuclei are labeled with Hoechst#33258 (blue) at 0.5 μg/mL. Co-labeling appears in blue-green/orange in the merged pictures. Note that VH411-S-Tag co-localizes with hLDLR and is internalized by CHO-hLDLR-EGFP cells. (B) Representative confocal photomicrographs of CHO-hLDLR-EGFP cells (green) incubated 1 hr at 37°C with a macromolecular complex (see scheme Fig 3B) resulting from the co-incubation and interaction of 10 μM VH411-S-Tag peptide or VH411Sc-S-Tag peptide with the anti-S-Tag antibody (1/200) and the Alexafluor 647-conjugated secondary antibody (1/800) (cyan). Cells were concomitantly exposed to 10 μg/mL of DiI-LDL (red). Cell nuclei are labeled with Hoechst#33258 (blue). Co-labeling appears in blue-green/orange in the merged pictures.

    Article Snippet: Cells were washed with PBS and incubated for 1 hr with primary antibodies directed against the hLDLR (rabbit, 1/100, Acris, Herford, Germany), against M13 bacteriophage (mouse, 1/3000, Sigma-Aldrich), against the S-Tag (goat, 1/250, Abcam) or against the Fc (mouse, 1/100, Jackson ImmunoResearch), followed by 3 washes with PBS and incubation with Alexa fluor 488, 594 or 647 anti-rabbit, anti-mouse or anti-goat secondary antibodies (1/800, Invitrogen).

    Techniques: Binding Assay, Incubation, Labeling

    Inhibition of the nuclear translocation of NF-κB upon GANT-61 treatment Immunofluorescence analysis was performed on MCF-7 and SK-BR-3 breast cancer cells after treatment with GANT-61 (10 μM) or vehicle alone for 24 hours. After the treatment, the cells were fixed, incubated with an anti-NF-κB antibody, washed, and labeled with an Alexa Fluor-488-conjugated goat anti-mouse IgG antibody. The nuclei were counterstained with Hoechst. Original magnification, 400x.

    Journal: Oncotarget

    Article Title: In vitro and in vivo inhibition of breast cancer cell growth by targeting the Hedgehog/GLI pathway with SMO (GDC-0449) or GLI (GANT-61) inhibitors

    doi: 10.18632/oncotarget.7062

    Figure Lengend Snippet: Inhibition of the nuclear translocation of NF-κB upon GANT-61 treatment Immunofluorescence analysis was performed on MCF-7 and SK-BR-3 breast cancer cells after treatment with GANT-61 (10 μM) or vehicle alone for 24 hours. After the treatment, the cells were fixed, incubated with an anti-NF-κB antibody, washed, and labeled with an Alexa Fluor-488-conjugated goat anti-mouse IgG antibody. The nuclei were counterstained with Hoechst. Original magnification, 400x.

    Article Snippet: Goat anti-rabbit IgG Alexa fluor-594-conjugated and goat anti-mouse IgG Alexa fluor-488-conjugated secondary antibody came from Invitrogen (Milano, Italy).

    Techniques: Inhibition, Translocation Assay, Immunofluorescence, Incubation, Labeling