goat anti mouse igg h l highly cross adsorbed secondary antibody  (Thermo Fisher)


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    Name:
    Goat anti Rabbit IgG H L Highly Cross Adsorbed Secondary Antibody
    Description:
    Goat anti Rabbit IgG H L Highly Cross Adsorbed Secondary Antibody for IF ICC Flow
    Catalog Number:
    A11034
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Antibodies and Secondary Detection|Cell Analysis|Secondary Detection
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    Structured Review

    Thermo Fisher goat anti mouse igg h l highly cross adsorbed secondary antibody
    Goat anti Rabbit IgG H L Highly Cross Adsorbed Secondary Antibody for IF ICC Flow
    https://www.bioz.com/result/goat anti mouse igg h l highly cross adsorbed secondary antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg h l highly cross adsorbed secondary antibody - by Bioz Stars, 2021-04
    97/100 stars

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    other:

    Article Title: Common cytotoxic chemotherapeutics induce epithelial-mesenchymal transition (EMT) downstream of ER stress
    Article Snippet: Alexa Fluor 488 goat anti-rabbit IgG #A11034 and Alexa Fluor 546 goat anti-rabbit IgG #A11010 (Molecular Probes, Invitrogen detection technologies, Eugene, OR.

    Incubation:

    Article Title: Formation and spreading of TDP-43 aggregates in cultured neuronal and glial cells demonstrated by time-lapse imaging
    Article Snippet: Immunocytochemistry Cells were fixed with 4% paraformaldehyde in phosphate buffered saline (PBS), permeabilized with 100% methanol, washed with PBS, and immunostained overnight at 4°C with the following primary antibodies at 1:200 dilutions; mouse monoclonal TuJ1 (R & D systems, Minneapolis, MN, USA), rabbit anti-phosphoTDP-43 (pSer409/410) (Cosmo Bio), rabbit anti-TDP-43 C-terminus (Abcam, Cambridge, MA, USA), rabbit anti-GFP (Abcam), rabbit anti-GFAP (DAKO, Glostrup, Denmark), rabbit anti-ubiquitin (DAKO) and rabbit anti-CNP (Cell Signaling Technology, Danvers, MA, USA). .. The cells were then incubated with Alexa Fluor 488- or 633-conjugated goat anti-rabbit or anti-mouse antibodies (Thermo Fisher) at 1:400 dilutions for 45 min at room temperature, followed by incubation for 15 min with 2 μg/mL Hoechst 33342 (Thermo Fisher). ..

    Article Title: Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway
    Article Snippet: 2.9.2 Immunofluorescence analyses Specimens were incubated with a blocking solution (1%BSA [Bovine Serum Albumin, Cat. No. A9647, Sigma-Aldrich], 0.5% Triton X-100 [Cat. No. T8787, Sigma-Aldrich] and 10% normal goat serum [Cat. No. S26-M, Sigma-Aldrich] in PBS 0.1 M), thereafter slices were incubated overnight at 4 °C with a solution containing anti-GSH (Cat. No. 19534, Abcam, Cambridge, MA, USA) or anti-HO-1 (Cat. No. ADI-SPA-896D Stressgen, Victoria, Canada) and Nrf2 (Cat. No. 89443, Abcam) primary antibodies diluted 1:100 in PBS. .. After washing in PBS, samples were incubated at room temperature for 2 h in labeled conjugated goat anti-rabbit (HO-1) (Alexa Fuor 488 Cat. No. A-11034, ThermoFischer, Waltham, MA, USA.) or donkey anti-mouse (GSH, Nrf2) secondary antibody (Alexa Fluor 488, Cat. No. A- 21202; or Alexa Fluor 546, IgG, Cat. No. A 10036, ThermoFischer) diluted 1:400 in 0.1 M PBS and stained with DAPI stained (1:500 in 0.1 M PBS). .. 2.10 Transfer assay for the non-metabolizable D -glucose analogue 2-NBDG 2-(N-(7-nitrobenz-2-oxa-1,3-dia-zol-4-yl)amino)-2-deoxyglucose (2-NBDG, ThermoFisher Cat. No. N13195, MW = 342.3) is a fluorescent glucose analogue that has been used to monitor glucose uptake in live cells and in the sensory epithelium of the cochlea .

    Labeling:

    Article Title: Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway
    Article Snippet: 2.9.2 Immunofluorescence analyses Specimens were incubated with a blocking solution (1%BSA [Bovine Serum Albumin, Cat. No. A9647, Sigma-Aldrich], 0.5% Triton X-100 [Cat. No. T8787, Sigma-Aldrich] and 10% normal goat serum [Cat. No. S26-M, Sigma-Aldrich] in PBS 0.1 M), thereafter slices were incubated overnight at 4 °C with a solution containing anti-GSH (Cat. No. 19534, Abcam, Cambridge, MA, USA) or anti-HO-1 (Cat. No. ADI-SPA-896D Stressgen, Victoria, Canada) and Nrf2 (Cat. No. 89443, Abcam) primary antibodies diluted 1:100 in PBS. .. After washing in PBS, samples were incubated at room temperature for 2 h in labeled conjugated goat anti-rabbit (HO-1) (Alexa Fuor 488 Cat. No. A-11034, ThermoFischer, Waltham, MA, USA.) or donkey anti-mouse (GSH, Nrf2) secondary antibody (Alexa Fluor 488, Cat. No. A- 21202; or Alexa Fluor 546, IgG, Cat. No. A 10036, ThermoFischer) diluted 1:400 in 0.1 M PBS and stained with DAPI stained (1:500 in 0.1 M PBS). .. 2.10 Transfer assay for the non-metabolizable D -glucose analogue 2-NBDG 2-(N-(7-nitrobenz-2-oxa-1,3-dia-zol-4-yl)amino)-2-deoxyglucose (2-NBDG, ThermoFisher Cat. No. N13195, MW = 342.3) is a fluorescent glucose analogue that has been used to monitor glucose uptake in live cells and in the sensory epithelium of the cochlea .

    Staining:

    Article Title: Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway
    Article Snippet: 2.9.2 Immunofluorescence analyses Specimens were incubated with a blocking solution (1%BSA [Bovine Serum Albumin, Cat. No. A9647, Sigma-Aldrich], 0.5% Triton X-100 [Cat. No. T8787, Sigma-Aldrich] and 10% normal goat serum [Cat. No. S26-M, Sigma-Aldrich] in PBS 0.1 M), thereafter slices were incubated overnight at 4 °C with a solution containing anti-GSH (Cat. No. 19534, Abcam, Cambridge, MA, USA) or anti-HO-1 (Cat. No. ADI-SPA-896D Stressgen, Victoria, Canada) and Nrf2 (Cat. No. 89443, Abcam) primary antibodies diluted 1:100 in PBS. .. After washing in PBS, samples were incubated at room temperature for 2 h in labeled conjugated goat anti-rabbit (HO-1) (Alexa Fuor 488 Cat. No. A-11034, ThermoFischer, Waltham, MA, USA.) or donkey anti-mouse (GSH, Nrf2) secondary antibody (Alexa Fluor 488, Cat. No. A- 21202; or Alexa Fluor 546, IgG, Cat. No. A 10036, ThermoFischer) diluted 1:400 in 0.1 M PBS and stained with DAPI stained (1:500 in 0.1 M PBS). .. 2.10 Transfer assay for the non-metabolizable D -glucose analogue 2-NBDG 2-(N-(7-nitrobenz-2-oxa-1,3-dia-zol-4-yl)amino)-2-deoxyglucose (2-NBDG, ThermoFisher Cat. No. N13195, MW = 342.3) is a fluorescent glucose analogue that has been used to monitor glucose uptake in live cells and in the sensory epithelium of the cochlea .

    Transduction:

    Article Title: Role of lipid rafts in E-cadherin- and HGF-R/Met-mediated entry of Listeria monocytogenes into host cells
    Article Snippet: .. Reagents and antibodies Alexa Fluor 488 cholera toxin subunit B, Alexa Fluor 488–, 546–, and 647–conjugated phalloidin, Alexa Fluor 488– and 546–conjugated goat anti–rabbit and goat anti–mouse antibodies were purchased from Molecular Probes. mAbs anti–E-cadherin (clone HECD-1) was purchased from R & D systems, anti–α-catenin (clone 5) was purchased from Transduction Laboratories. ..

    Immunofluorescence:

    Article Title: Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes
    Article Snippet: .. Indirect immunofluorescence was performed using secondary antibodies conjugated with Alexa-647 anti-Mouse (Invitrogen, Cat. A-21236), Alexa-555 anti-Rat (Invitrogen, Cat. A-21434) and Alexa-488 anti-Rabbit (Invitrogen, Cat. A-11034). .. 10 mg/ml Hoechst 33342 stock solution was purchased from Life Technologies (Cat. H3570).

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  • 97
    Thermo Fisher goat anti mouse fitc
    US27 is found in close proximity to <t>CXCR4.</t> (A) Immunofluorescence microscopy of HEK293-US27 or HEK293-US28 cells stained with anti-FLAG (green) and anti-CXCR4 (red) followed by goat anti-mouse <t>FITC</t> and donkey anti-goat TRITC secondary antibodies, respectively. Nuclei were visualized with DAPI (blue). Bar, 10 μm. (B) HEK293-US27 or HEK293-US28 cells were stained with anti-FLAG and anti-CXCR4 followed by oligonucleotide-conjugated secondary antibodies and amplification using the DuoLink kit for the proximity ligation assay (PLA). Red PLA spots indicate receptors in close proximity. (C) NuFF cells were infected with AD169-GFP (MOI = 1, 72 hpi) and stained with antibodies against CXCR4 and US27 (left) or CXCR4 and US28 (right) followed by DuoLink PLA. Infected cells are shown in green, and red indicates PLA spots. Images were captured using a Zeiss LSM700 laser scanning confocal microscope, and representative images are shown. For PLA (B, C), images are maximum-intensity projections from z-stacks.
    Goat Anti Mouse Fitc, 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 mouse fitc/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
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    95
    Thermo Fisher cross adsorbed goat anti mouse igg conjugated to alexa fluor 568
    Confocal microscopy and adherence-invasion assays with pharmacologic inhibitors. Lipid raft-independent invasion of bronchial epithelial cells by NTHI. (A) Confocal microscopy. At 4 and 24 h postinoculation of H292 cells, NTHI does not colocalize with vesicles positive for the following markers of lipid rafts: caveolin-1 (shown at 24 h), flotillin-1 (shown at 24 h), and cholesterol (shown at 4 h). NTHI was labeled with anti-11P6H antibody conjugated to <t>Alexa</t> Fluor 488 (green). Caveolin-1 was labeled with anti-caveolin-1 antibody and goat anti-mouse antibody conjugated to Alexa <t>Fluor</t> 568 (red). Flotillin-1 was labeled with anti-flotillin-1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). Cholesterol was labeled with filipin (blue), and those samples were also labeled with anti-human secretory component antibody and donkey anti-goat antibody conjugated to Alexa Fluor 568 (red) to provide additional visualization of the plasma membrane. (B) Confocal microscopy. Filipin (FLP) and nystatin (NST) inhibit the internalization of fluorescently conjugated cholera toxin B subunit (white arrows), a known cargo of lipid raft-mediated endocytosis. H292 cells were pretreated with sRPMI containing inhibitor or inhibitor diluent, followed by addition of the cargo conjugate, incubation for 20 min on ice, and incubation for 2 h at 37°C. (C) Adherence-invasion assays. Filipin and nystatin do not inhibit invasion by NTHI. H292 cells were pretreated with inhibitor or inhibitor diluent for 2 h, followed by a 4-h infection conducted according to the adherence-invasion assay protocol. Data are normalized to samples in the absence of inhibitor. Error bars represent standard errors of the means of three independent experiments. A paired Student t test was used to calculate statistical significance. *, P ≤ 0.05; **, P ≤ 0.005.
    Cross Adsorbed Goat Anti Mouse Igg Conjugated To Alexa Fluor 568, 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
    https://www.bioz.com/result/cross adsorbed goat anti mouse igg conjugated to alexa fluor 568/product/Thermo Fisher
    Average 95 stars, based on 1 article reviews
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    94
    Thermo Fisher peroxidase conjugated goat anti mouse immunoglobulins
    Determination by Western blotting of the specificity of MAbs with regard to K88ab, K88ac, and K88ad adhesins. K88ab, K88ac, and K88ad adhesin antigens were separated by SDS-PAGE and transferred to nitrocellulose membranes. These membranes were probed with MAb 36/41 or 221/38 or BSA, followed by incubation with <t>peroxidase-conjugated</t> <t>goat</t> <t>anti-mouse</t> IgG as described in Materials and Methods. Molecular mass standards are indicated on the left. MAb 36/41 binds to K88ac fimbriae specifically, and MAb 221/38 binds to all three variants of K88 fimbriae.
    Peroxidase Conjugated Goat Anti Mouse Immunoglobulins, 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
    https://www.bioz.com/result/peroxidase conjugated goat anti mouse immunoglobulins/product/Thermo Fisher
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    Image Search Results


    US27 is found in close proximity to CXCR4. (A) Immunofluorescence microscopy of HEK293-US27 or HEK293-US28 cells stained with anti-FLAG (green) and anti-CXCR4 (red) followed by goat anti-mouse FITC and donkey anti-goat TRITC secondary antibodies, respectively. Nuclei were visualized with DAPI (blue). Bar, 10 μm. (B) HEK293-US27 or HEK293-US28 cells were stained with anti-FLAG and anti-CXCR4 followed by oligonucleotide-conjugated secondary antibodies and amplification using the DuoLink kit for the proximity ligation assay (PLA). Red PLA spots indicate receptors in close proximity. (C) NuFF cells were infected with AD169-GFP (MOI = 1, 72 hpi) and stained with antibodies against CXCR4 and US27 (left) or CXCR4 and US28 (right) followed by DuoLink PLA. Infected cells are shown in green, and red indicates PLA spots. Images were captured using a Zeiss LSM700 laser scanning confocal microscope, and representative images are shown. For PLA (B, C), images are maximum-intensity projections from z-stacks.

    Journal: Journal of Virology

    Article Title: Human Cytomegalovirus UL111A and US27 Gene Products Enhance the CXCL12/CXCR4 Signaling Axis via Distinct Mechanisms

    doi: 10.1128/JVI.01981-17

    Figure Lengend Snippet: US27 is found in close proximity to CXCR4. (A) Immunofluorescence microscopy of HEK293-US27 or HEK293-US28 cells stained with anti-FLAG (green) and anti-CXCR4 (red) followed by goat anti-mouse FITC and donkey anti-goat TRITC secondary antibodies, respectively. Nuclei were visualized with DAPI (blue). Bar, 10 μm. (B) HEK293-US27 or HEK293-US28 cells were stained with anti-FLAG and anti-CXCR4 followed by oligonucleotide-conjugated secondary antibodies and amplification using the DuoLink kit for the proximity ligation assay (PLA). Red PLA spots indicate receptors in close proximity. (C) NuFF cells were infected with AD169-GFP (MOI = 1, 72 hpi) and stained with antibodies against CXCR4 and US27 (left) or CXCR4 and US28 (right) followed by DuoLink PLA. Infected cells are shown in green, and red indicates PLA spots. Images were captured using a Zeiss LSM700 laser scanning confocal microscope, and representative images are shown. For PLA (B, C), images are maximum-intensity projections from z-stacks.

    Article Snippet: For standard immunofluorescence (IF), cells were costained with anti-CXCR4 goat polyclonal (1:100) and anti-FLAG mouse monoclonal antibody (1:500) followed by goat anti-mouse FITC and donkey anti-goat tetramethylrhodamine (TRTIC; 1:250), and then Prolong Gold mounting medium with DAPI (4′,6-diamidino-2-phenylindole; Invitrogen) was added.

    Techniques: Immunofluorescence, Microscopy, Staining, Amplification, Proximity Ligation Assay, Infection

    cmvIL-10 enhances CXCR4 signaling in HCMV-infected cells. (A) NuFF cells were infected with TB40/E- mCherry (MOI = 1). Total RNA was harvested from mock- or HCMV-infected NuFFs at 72 h postinfection (hpi) and reverse transcribed, and UL123 (IE1) or β-actin genes were amplified. The resulting bands were visualized via agarose gel electrophoresis. (B) Mock- or HCMV-infected NuFF cells were stained with IL-10R-FITC or isotype control antibody and analyzed by flow cytometry. (C) Flow cytometric comparison of mock- or HCMV-infected NuFF cells for mCherry fluorescence as a measure of infection (left) or IL-10R stained as described above (right). (D) Calcium response was evaluated in Fluo-4 AM-loaded mock- or HCMV-infected cells stimulated with 100 ng/ml CXCL12 ± 100 ng/ml cmvIL-10 at 72 hpi. Relative fluorescence units (RFU) were measured by flow cytometry; the arrow indicates stimulus addition. (E) Transwell migration of mock-infected (gray bars) or HCMV-infected (black bars) NuFF cells toward CXCL12 in the lower chamber ± 100 ng/ml cmvIL-10 (striped bars, mock infected; white bars, HCMV infected) was measured at 72 hpi. Error bars, standard error for 3 replicates; *, P

    Journal: Journal of Virology

    Article Title: Human Cytomegalovirus UL111A and US27 Gene Products Enhance the CXCL12/CXCR4 Signaling Axis via Distinct Mechanisms

    doi: 10.1128/JVI.01981-17

    Figure Lengend Snippet: cmvIL-10 enhances CXCR4 signaling in HCMV-infected cells. (A) NuFF cells were infected with TB40/E- mCherry (MOI = 1). Total RNA was harvested from mock- or HCMV-infected NuFFs at 72 h postinfection (hpi) and reverse transcribed, and UL123 (IE1) or β-actin genes were amplified. The resulting bands were visualized via agarose gel electrophoresis. (B) Mock- or HCMV-infected NuFF cells were stained with IL-10R-FITC or isotype control antibody and analyzed by flow cytometry. (C) Flow cytometric comparison of mock- or HCMV-infected NuFF cells for mCherry fluorescence as a measure of infection (left) or IL-10R stained as described above (right). (D) Calcium response was evaluated in Fluo-4 AM-loaded mock- or HCMV-infected cells stimulated with 100 ng/ml CXCL12 ± 100 ng/ml cmvIL-10 at 72 hpi. Relative fluorescence units (RFU) were measured by flow cytometry; the arrow indicates stimulus addition. (E) Transwell migration of mock-infected (gray bars) or HCMV-infected (black bars) NuFF cells toward CXCL12 in the lower chamber ± 100 ng/ml cmvIL-10 (striped bars, mock infected; white bars, HCMV infected) was measured at 72 hpi. Error bars, standard error for 3 replicates; *, P

    Article Snippet: For standard immunofluorescence (IF), cells were costained with anti-CXCR4 goat polyclonal (1:100) and anti-FLAG mouse monoclonal antibody (1:500) followed by goat anti-mouse FITC and donkey anti-goat tetramethylrhodamine (TRTIC; 1:250), and then Prolong Gold mounting medium with DAPI (4′,6-diamidino-2-phenylindole; Invitrogen) was added.

    Techniques: Infection, Amplification, Agarose Gel Electrophoresis, Staining, Flow Cytometry, Cytometry, Fluorescence, Migration

    Confocal microscopy and adherence-invasion assays with pharmacologic inhibitors. Lipid raft-independent invasion of bronchial epithelial cells by NTHI. (A) Confocal microscopy. At 4 and 24 h postinoculation of H292 cells, NTHI does not colocalize with vesicles positive for the following markers of lipid rafts: caveolin-1 (shown at 24 h), flotillin-1 (shown at 24 h), and cholesterol (shown at 4 h). NTHI was labeled with anti-11P6H antibody conjugated to Alexa Fluor 488 (green). Caveolin-1 was labeled with anti-caveolin-1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). Flotillin-1 was labeled with anti-flotillin-1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). Cholesterol was labeled with filipin (blue), and those samples were also labeled with anti-human secretory component antibody and donkey anti-goat antibody conjugated to Alexa Fluor 568 (red) to provide additional visualization of the plasma membrane. (B) Confocal microscopy. Filipin (FLP) and nystatin (NST) inhibit the internalization of fluorescently conjugated cholera toxin B subunit (white arrows), a known cargo of lipid raft-mediated endocytosis. H292 cells were pretreated with sRPMI containing inhibitor or inhibitor diluent, followed by addition of the cargo conjugate, incubation for 20 min on ice, and incubation for 2 h at 37°C. (C) Adherence-invasion assays. Filipin and nystatin do not inhibit invasion by NTHI. H292 cells were pretreated with inhibitor or inhibitor diluent for 2 h, followed by a 4-h infection conducted according to the adherence-invasion assay protocol. Data are normalized to samples in the absence of inhibitor. Error bars represent standard errors of the means of three independent experiments. A paired Student t test was used to calculate statistical significance. *, P ≤ 0.05; **, P ≤ 0.005.

    Journal: Infection and Immunity

    Article Title: Internalization and Trafficking of Nontypeable Haemophilus influenzae in Human Respiratory Epithelial Cells and Roles of IgA1 Proteases for Optimal Invasion and Persistence

    doi: 10.1128/IAI.00864-13

    Figure Lengend Snippet: Confocal microscopy and adherence-invasion assays with pharmacologic inhibitors. Lipid raft-independent invasion of bronchial epithelial cells by NTHI. (A) Confocal microscopy. At 4 and 24 h postinoculation of H292 cells, NTHI does not colocalize with vesicles positive for the following markers of lipid rafts: caveolin-1 (shown at 24 h), flotillin-1 (shown at 24 h), and cholesterol (shown at 4 h). NTHI was labeled with anti-11P6H antibody conjugated to Alexa Fluor 488 (green). Caveolin-1 was labeled with anti-caveolin-1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). Flotillin-1 was labeled with anti-flotillin-1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). Cholesterol was labeled with filipin (blue), and those samples were also labeled with anti-human secretory component antibody and donkey anti-goat antibody conjugated to Alexa Fluor 568 (red) to provide additional visualization of the plasma membrane. (B) Confocal microscopy. Filipin (FLP) and nystatin (NST) inhibit the internalization of fluorescently conjugated cholera toxin B subunit (white arrows), a known cargo of lipid raft-mediated endocytosis. H292 cells were pretreated with sRPMI containing inhibitor or inhibitor diluent, followed by addition of the cargo conjugate, incubation for 20 min on ice, and incubation for 2 h at 37°C. (C) Adherence-invasion assays. Filipin and nystatin do not inhibit invasion by NTHI. H292 cells were pretreated with inhibitor or inhibitor diluent for 2 h, followed by a 4-h infection conducted according to the adherence-invasion assay protocol. Data are normalized to samples in the absence of inhibitor. Error bars represent standard errors of the means of three independent experiments. A paired Student t test was used to calculate statistical significance. *, P ≤ 0.05; **, P ≤ 0.005.

    Article Snippet: Host cell components were visualized using filipin III fluorescent dye (final concentration of 333 μg/ml; Cayman Chemical), primary monoclonal antibodies including mouse anti-caveolin-1 antibody (1:50; BD), mouse anti-early endosomal antigen 1 (EEA-1) antibody (1:200; BD), mouse anti-flotillin-1 antibody (1:20; BD), mouse anti-human CD107a antibody (LAMP1) (1:200; BD), and goat anti-human secretory component antibody (1:200; Sigma), and secondary antibodies including highly cross-adsorbed goat anti-mouse IgG conjugated to Alexa Fluor 568 (Life Technologies, 1:200) and donkey anti-goat IgG conjugated to Alexa Fluor 568 (Life Technologies, 1:200).

    Techniques: Confocal Microscopy, Labeling, Incubation, Infection, Invasion Assay

    Confocal microscopy and survival assay with pharmacologic inhibitor of lysosome acidification. NTHI traffics to early endosomes and lysosomes and is killed in lysosomes. IgA1 proteases are required for optimal survival in lysosomes. (A) Confocal microscopy. At 4 and 24 h postinoculation of H292 cells, NTHI is found within vesicles positive for EEA1 (shown at 4 h) and within vesicles positive for LAMP1 (shown at 24 h). NTHI was labeled using anti-11P6H antibody conjugated to Alexa Fluor 488 (green). EEA1 was labeled using anti-EEA1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). LAMP1 was labeled using anti-human LAMP1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). (B) Survival assay in the presence and absence of concanamycin A (CMA). H292 cells were infected for 16 h, treated with gentamicin for 1 h, and treated with concanamycin or diluent for 3 h. Survival data are normalized to samples treated with diluent. Error bars represent standard errors of the means of three independent experiments. A paired Student t test was used to calculate statistical significance. *, P ≤ 0.05; **, P ≤ 0.005.

    Journal: Infection and Immunity

    Article Title: Internalization and Trafficking of Nontypeable Haemophilus influenzae in Human Respiratory Epithelial Cells and Roles of IgA1 Proteases for Optimal Invasion and Persistence

    doi: 10.1128/IAI.00864-13

    Figure Lengend Snippet: Confocal microscopy and survival assay with pharmacologic inhibitor of lysosome acidification. NTHI traffics to early endosomes and lysosomes and is killed in lysosomes. IgA1 proteases are required for optimal survival in lysosomes. (A) Confocal microscopy. At 4 and 24 h postinoculation of H292 cells, NTHI is found within vesicles positive for EEA1 (shown at 4 h) and within vesicles positive for LAMP1 (shown at 24 h). NTHI was labeled using anti-11P6H antibody conjugated to Alexa Fluor 488 (green). EEA1 was labeled using anti-EEA1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). LAMP1 was labeled using anti-human LAMP1 antibody and goat anti-mouse antibody conjugated to Alexa Fluor 568 (red). (B) Survival assay in the presence and absence of concanamycin A (CMA). H292 cells were infected for 16 h, treated with gentamicin for 1 h, and treated with concanamycin or diluent for 3 h. Survival data are normalized to samples treated with diluent. Error bars represent standard errors of the means of three independent experiments. A paired Student t test was used to calculate statistical significance. *, P ≤ 0.05; **, P ≤ 0.005.

    Article Snippet: Host cell components were visualized using filipin III fluorescent dye (final concentration of 333 μg/ml; Cayman Chemical), primary monoclonal antibodies including mouse anti-caveolin-1 antibody (1:50; BD), mouse anti-early endosomal antigen 1 (EEA-1) antibody (1:200; BD), mouse anti-flotillin-1 antibody (1:20; BD), mouse anti-human CD107a antibody (LAMP1) (1:200; BD), and goat anti-human secretory component antibody (1:200; Sigma), and secondary antibodies including highly cross-adsorbed goat anti-mouse IgG conjugated to Alexa Fluor 568 (Life Technologies, 1:200) and donkey anti-goat IgG conjugated to Alexa Fluor 568 (Life Technologies, 1:200).

    Techniques: Confocal Microscopy, Clonogenic Cell Survival Assay, Labeling, Infection

    Determination by Western blotting of the specificity of MAbs with regard to K88ab, K88ac, and K88ad adhesins. K88ab, K88ac, and K88ad adhesin antigens were separated by SDS-PAGE and transferred to nitrocellulose membranes. These membranes were probed with MAb 36/41 or 221/38 or BSA, followed by incubation with peroxidase-conjugated goat anti-mouse IgG as described in Materials and Methods. Molecular mass standards are indicated on the left. MAb 36/41 binds to K88ac fimbriae specifically, and MAb 221/38 binds to all three variants of K88 fimbriae.

    Journal: Infection and Immunity

    Article Title: Inhibition of Adhesion of Escherichia coli K88ac Fimbria to Its Receptor, Intestinal Mucin-Type Glycoproteins, by a Monoclonal Antibody Directed against a Variable Domain of the Fimbria

    doi:

    Figure Lengend Snippet: Determination by Western blotting of the specificity of MAbs with regard to K88ab, K88ac, and K88ad adhesins. K88ab, K88ac, and K88ad adhesin antigens were separated by SDS-PAGE and transferred to nitrocellulose membranes. These membranes were probed with MAb 36/41 or 221/38 or BSA, followed by incubation with peroxidase-conjugated goat anti-mouse IgG as described in Materials and Methods. Molecular mass standards are indicated on the left. MAb 36/41 binds to K88ac fimbriae specifically, and MAb 221/38 binds to all three variants of K88 fimbriae.

    Article Snippet: The plates were then washed as before, peroxidase-conjugated goat anti-mouse immunoglobulins (Pierce, Rockford, Ill.) were added, and the plates were again incubated.

    Techniques: Western Blot, SDS Page, Incubation