alexa fluor 594 conjugated concanavalin a  (Thermo Fisher)


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    Name:
    Concanavalin A Alexa Fluor 594 Conjugate
    Description:
    Concanavalin A Con A is one of the most widely used lectins in cell biology Our Alexa Fluor 594 conjugate of Con A exhibits the bright red fluorescence of the Alexa Fluor 594 dye absorption emission maxima 590 617 nm Alexa Fluor 594 Con A selectively binds to a mannopyranosyl and a glucopyranosyl residues
    Catalog Number:
    c11253
    Price:
    None
    Category:
    Labeling Detection Products
    Applications:
    Cell Analysis|Cellular Imaging|Flow Cytometry Antibodies & Secondary Detection|General Cell Tracing|Immunofluorescence (IF)|Immunofluorescence Staining & Detection|Microbial Tracking|Flow Cytometry|Cell Tracing & Tracking
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    Structured Review

    Thermo Fisher alexa fluor 594 conjugated concanavalin a
    PM Localization of HTLV-1 MA GagLZ, MLV MA GagLZ, and HERV-K MA GagLZ persists upon 5ptaseIV overexpression, unlike that of HIV-1 GagLZ and RSV MA GagLZ. (A) Schematic illustrations of HIV-1 Gag, HIV-1 GagLZ, RSV MA GagLZ, HTLV-1 MA GagLZ, MLV MA GagLZ, and HERV-K MA GagLZ are shown. The first amino acid, methionine, is included in the numbering of MA residues, although the methionine is removed upon N-terminal myristylation. All Gag proteins were expressed from the same CMV-promoter-driven vector backbone. (B) HeLa cells expressing YFP-tagged HIV-1 GagLZ, RSV MA GagLZ, HTLV-1 MA GagLZ, MLV MA GagLZ, or HERV-K MA GagLZ, along with Myc-tagged full-length (FL) 5ptaseIV or the Δ1 derivative, were stained with ConA labeled with <t>Alexa</t> <t>Fluor</t> 594 (not shown), immunostained with mouse monoclonal anti-Myc antibody and anti-mouse IgG conjugated with Alexa Fluor 647 (anti-Myc), and analyzed using a confocal fluorescence microscope. Note that overexpression of 5ptaseIV FL induced mislocalization of HIV-1 GagLZ and RSV MA GagLZ to the cytosol and abolished PM localization. In contrast, 5ptaseIV FL overexpression did not drastically alter localization of HTLV-1 MA GagLZ and MLV MA GagLZ to the PM and intracellular compartments. Overexpression of 5ptaseIV increased the localization of HERV-K MA GagLZ to intracellular compartments, but PM localization persisted regardless of the intracellular localization. (C) HeLa cells expressing YFP-tagged HIV-1 Gag, HTMA Gag, and HIV-1 (1GA) Gag were stained with ConA labeled with Alexa Fluor 594. Cells were then fixed and analyzed using a confocal fluorescence microscope. (D) Pearson’s correlation coefficients (PCC) for colocalization of Gag-YFP or GagLZ-YFP with ConA were calculated and are shown as means ± SEM. Twenty to fifty cells were analyzed per condition. **, P
    Concanavalin A Con A is one of the most widely used lectins in cell biology Our Alexa Fluor 594 conjugate of Con A exhibits the bright red fluorescence of the Alexa Fluor 594 dye absorption emission maxima 590 617 nm Alexa Fluor 594 Con A selectively binds to a mannopyranosyl and a glucopyranosyl residues
    https://www.bioz.com/result/alexa fluor 594 conjugated concanavalin a/product/Thermo Fisher
    Average 93 stars, based on 1 article reviews
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    alexa fluor 594 conjugated concanavalin a - by Bioz Stars, 2021-03
    93/100 stars

    Images

    1) Product Images from "Membrane Binding and Subcellular Localization of Retroviral Gag Proteins Are Differentially Regulated by MA Interactions with Phosphatidylinositol-(4,5)-Bisphosphate and RNA"

    Article Title: Membrane Binding and Subcellular Localization of Retroviral Gag Proteins Are Differentially Regulated by MA Interactions with Phosphatidylinositol-(4,5)-Bisphosphate and RNA

    Journal: mBio

    doi: 10.1128/mBio.02202-14

    PM Localization of HTLV-1 MA GagLZ, MLV MA GagLZ, and HERV-K MA GagLZ persists upon 5ptaseIV overexpression, unlike that of HIV-1 GagLZ and RSV MA GagLZ. (A) Schematic illustrations of HIV-1 Gag, HIV-1 GagLZ, RSV MA GagLZ, HTLV-1 MA GagLZ, MLV MA GagLZ, and HERV-K MA GagLZ are shown. The first amino acid, methionine, is included in the numbering of MA residues, although the methionine is removed upon N-terminal myristylation. All Gag proteins were expressed from the same CMV-promoter-driven vector backbone. (B) HeLa cells expressing YFP-tagged HIV-1 GagLZ, RSV MA GagLZ, HTLV-1 MA GagLZ, MLV MA GagLZ, or HERV-K MA GagLZ, along with Myc-tagged full-length (FL) 5ptaseIV or the Δ1 derivative, were stained with ConA labeled with Alexa Fluor 594 (not shown), immunostained with mouse monoclonal anti-Myc antibody and anti-mouse IgG conjugated with Alexa Fluor 647 (anti-Myc), and analyzed using a confocal fluorescence microscope. Note that overexpression of 5ptaseIV FL induced mislocalization of HIV-1 GagLZ and RSV MA GagLZ to the cytosol and abolished PM localization. In contrast, 5ptaseIV FL overexpression did not drastically alter localization of HTLV-1 MA GagLZ and MLV MA GagLZ to the PM and intracellular compartments. Overexpression of 5ptaseIV increased the localization of HERV-K MA GagLZ to intracellular compartments, but PM localization persisted regardless of the intracellular localization. (C) HeLa cells expressing YFP-tagged HIV-1 Gag, HTMA Gag, and HIV-1 (1GA) Gag were stained with ConA labeled with Alexa Fluor 594. Cells were then fixed and analyzed using a confocal fluorescence microscope. (D) Pearson’s correlation coefficients (PCC) for colocalization of Gag-YFP or GagLZ-YFP with ConA were calculated and are shown as means ± SEM. Twenty to fifty cells were analyzed per condition. **, P
    Figure Legend Snippet: PM Localization of HTLV-1 MA GagLZ, MLV MA GagLZ, and HERV-K MA GagLZ persists upon 5ptaseIV overexpression, unlike that of HIV-1 GagLZ and RSV MA GagLZ. (A) Schematic illustrations of HIV-1 Gag, HIV-1 GagLZ, RSV MA GagLZ, HTLV-1 MA GagLZ, MLV MA GagLZ, and HERV-K MA GagLZ are shown. The first amino acid, methionine, is included in the numbering of MA residues, although the methionine is removed upon N-terminal myristylation. All Gag proteins were expressed from the same CMV-promoter-driven vector backbone. (B) HeLa cells expressing YFP-tagged HIV-1 GagLZ, RSV MA GagLZ, HTLV-1 MA GagLZ, MLV MA GagLZ, or HERV-K MA GagLZ, along with Myc-tagged full-length (FL) 5ptaseIV or the Δ1 derivative, were stained with ConA labeled with Alexa Fluor 594 (not shown), immunostained with mouse monoclonal anti-Myc antibody and anti-mouse IgG conjugated with Alexa Fluor 647 (anti-Myc), and analyzed using a confocal fluorescence microscope. Note that overexpression of 5ptaseIV FL induced mislocalization of HIV-1 GagLZ and RSV MA GagLZ to the cytosol and abolished PM localization. In contrast, 5ptaseIV FL overexpression did not drastically alter localization of HTLV-1 MA GagLZ and MLV MA GagLZ to the PM and intracellular compartments. Overexpression of 5ptaseIV increased the localization of HERV-K MA GagLZ to intracellular compartments, but PM localization persisted regardless of the intracellular localization. (C) HeLa cells expressing YFP-tagged HIV-1 Gag, HTMA Gag, and HIV-1 (1GA) Gag were stained with ConA labeled with Alexa Fluor 594. Cells were then fixed and analyzed using a confocal fluorescence microscope. (D) Pearson’s correlation coefficients (PCC) for colocalization of Gag-YFP or GagLZ-YFP with ConA were calculated and are shown as means ± SEM. Twenty to fifty cells were analyzed per condition. **, P

    Techniques Used: Over Expression, Plasmid Preparation, Expressing, Staining, Labeling, Fluorescence, Microscopy, Periodic Counter-current Chromatography

    2) Product Images from "Disease-causing Mutation in PKR2 Receptor Reveals a Critical Role of Positive Charges in the Second Intracellular Loop for G-protein Coupling and Receptor Trafficking"

    Article Title: Disease-causing Mutation in PKR2 Receptor Reveals a Critical Role of Positive Charges in the Second Intracellular Loop for G-protein Coupling and Receptor Trafficking

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.223784

    The effect of R164Q mutation on the signal transduction and cell surface expression of PKR2. A , R164Q mutation disrupted the signal transduction of PKR2 as assayed with an aequorin-based assay. B , both WT and R164Q-PKR2 were presented on the cell surface. The Alexa Fluor 594-conjugated concanavalin A was used to label the membrane. The fluorescence was monitored with a confocal microscope. Scale bar: 10 μm.
    Figure Legend Snippet: The effect of R164Q mutation on the signal transduction and cell surface expression of PKR2. A , R164Q mutation disrupted the signal transduction of PKR2 as assayed with an aequorin-based assay. B , both WT and R164Q-PKR2 were presented on the cell surface. The Alexa Fluor 594-conjugated concanavalin A was used to label the membrane. The fluorescence was monitored with a confocal microscope. Scale bar: 10 μm.

    Techniques Used: Mutagenesis, Transduction, Expressing, Fluorescence, Microscopy

    3) Product Images from "Relationships between MA-RNA Binding in Cells and Suppression of HIV-1 Gag Mislocalization to Intracellular Membranes"

    Article Title: Relationships between MA-RNA Binding in Cells and Suppression of HIV-1 Gag Mislocalization to Intracellular Membranes

    Journal: Journal of Virology

    doi: 10.1128/JVI.00756-19

    Gag derivatives containing KR substitutions in the MA-HBR do not show promiscuous localization in cells, unlike those with KT changes. (A and B) HeLa cells were transfected with full-length Gag-YFP (A) and delNC/Gag-YFP (B), which contain the WT MA sequence or 25/26KR, 25/26KT, 29/31KR, or 29/31KT. At 14 h posttransfection, cells were stained with Alexa Fluor 594-conjugated concanavalin A (ConA-AF594), fixed with 4% paraformaldehyde in PBS, and analyzed using a fluorescence microscope. Note that subcellular distributions of delNC/Gag-YFP constructs mirror those of the corresponding full-length Gag-YFP constructs for both WT and MA-HBR mutants. Forty-two to 85 cells were analyzed under each condition across 3 independent experiments. The localization patterns determined by epifluorescence microscopy were confirmed by confocal microscopy. (Top) Representative confocal images. The blue arrowhead indicates the Gag-YFP signal in the intracellular compartments (IC). (Bottom) Bar graphs representing percentages of cells showing the indicated patterns of subcellular distribution for each Gag-YFP construct. ConA-AF594 staining was used as a plasma membrane (PM) marker (not shown). (C, top) Images showing an example of Gag-YFP showing PM-specific localization and ConA-AF594 staining of the PM. Green arrowheads indicate overlap of the two signals at the cell surface in a merged image. (Bottom) Images showing an example of Gag-YFP exhibiting PM and IC localization. The blue arrowhead indicates the Gag-YFP signal in the intracellular compartments. (D) HeLa cells were transfected with full-length WT Gag-YFP or 29/31KA Gag-YFP. At 16 h posttransfection, cells were stained with ConA-AF594, fixed with 4% paraformaldehyde in PBS, and analyzed using a fluorescence microscope. A total of 45 to 62 cells in 3 independent experiments were analyzed. The localization patterns determined by epifluorescence microscopy were confirmed by confocal microscopy. (Top) Representative confocal images. The blue arrowhead indicates the Gag-YFP signal in the intracellular compartments. (Bottom) Bar graphs representing percentages of cell populations as described above.
    Figure Legend Snippet: Gag derivatives containing KR substitutions in the MA-HBR do not show promiscuous localization in cells, unlike those with KT changes. (A and B) HeLa cells were transfected with full-length Gag-YFP (A) and delNC/Gag-YFP (B), which contain the WT MA sequence or 25/26KR, 25/26KT, 29/31KR, or 29/31KT. At 14 h posttransfection, cells were stained with Alexa Fluor 594-conjugated concanavalin A (ConA-AF594), fixed with 4% paraformaldehyde in PBS, and analyzed using a fluorescence microscope. Note that subcellular distributions of delNC/Gag-YFP constructs mirror those of the corresponding full-length Gag-YFP constructs for both WT and MA-HBR mutants. Forty-two to 85 cells were analyzed under each condition across 3 independent experiments. The localization patterns determined by epifluorescence microscopy were confirmed by confocal microscopy. (Top) Representative confocal images. The blue arrowhead indicates the Gag-YFP signal in the intracellular compartments (IC). (Bottom) Bar graphs representing percentages of cells showing the indicated patterns of subcellular distribution for each Gag-YFP construct. ConA-AF594 staining was used as a plasma membrane (PM) marker (not shown). (C, top) Images showing an example of Gag-YFP showing PM-specific localization and ConA-AF594 staining of the PM. Green arrowheads indicate overlap of the two signals at the cell surface in a merged image. (Bottom) Images showing an example of Gag-YFP exhibiting PM and IC localization. The blue arrowhead indicates the Gag-YFP signal in the intracellular compartments. (D) HeLa cells were transfected with full-length WT Gag-YFP or 29/31KA Gag-YFP. At 16 h posttransfection, cells were stained with ConA-AF594, fixed with 4% paraformaldehyde in PBS, and analyzed using a fluorescence microscope. A total of 45 to 62 cells in 3 independent experiments were analyzed. The localization patterns determined by epifluorescence microscopy were confirmed by confocal microscopy. (Top) Representative confocal images. The blue arrowhead indicates the Gag-YFP signal in the intracellular compartments. (Bottom) Bar graphs representing percentages of cell populations as described above.

    Techniques Used: Transfection, Sequencing, Staining, Fluorescence, Microscopy, Construct, Epifluorescence Microscopy, Confocal Microscopy, Marker

    4) Product Images from "Molecular Determinants Directing HIV-1 Gag Assembly to Virus-Containing Compartments in Primary Macrophages"

    Article Title: Molecular Determinants Directing HIV-1 Gag Assembly to Virus-Containing Compartments in Primary Macrophages

    Journal: Journal of Virology

    doi: 10.1128/JVI.01004-16

    HIV-1 Gag failed to localize to VCCs upon 5ptaseIV overexpression. (A) MDMs were infected with pseudotyped HIV-1 encoding Gag-YFP along with 5ptaseIV (FL) or 5ptaseIV Δ1. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594 (a PM marker), fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. (B) Pearson's correlation coefficients for colocalization of Gag-YFP with CD81 are shown as means and standard errors of the means (SEM). Twenty to 30 cells were analyzed per condition. **, P
    Figure Legend Snippet: HIV-1 Gag failed to localize to VCCs upon 5ptaseIV overexpression. (A) MDMs were infected with pseudotyped HIV-1 encoding Gag-YFP along with 5ptaseIV (FL) or 5ptaseIV Δ1. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594 (a PM marker), fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. (B) Pearson's correlation coefficients for colocalization of Gag-YFP with CD81 are shown as means and standard errors of the means (SEM). Twenty to 30 cells were analyzed per condition. **, P

    Techniques Used: Over Expression, Infection, Staining, Labeling, Marker, Microscopy

    The myristate moiety but not the intact HIV-1 MA sequence is required for VCC localization. (A) MDMs were infected with pseudotyped HIV-1 encoding WT Gag-YFP or Gag-YFP derivatives containing the indicated MA substitutions. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594, fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. (B) Pearson's correlation coefficients for colocalization of Gag-YFP with CD81 are shown as means and SEM. Twenty to 30 cells were analyzed per condition. *, P
    Figure Legend Snippet: The myristate moiety but not the intact HIV-1 MA sequence is required for VCC localization. (A) MDMs were infected with pseudotyped HIV-1 encoding WT Gag-YFP or Gag-YFP derivatives containing the indicated MA substitutions. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594, fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. (B) Pearson's correlation coefficients for colocalization of Gag-YFP with CD81 are shown as means and SEM. Twenty to 30 cells were analyzed per condition. *, P

    Techniques Used: Sequencing, Infection, Staining, Labeling, Microscopy

    Heterologous membrane binding sequences can replace HIV-1 MA without affecting VCC localization. (A) Schematic illustrations of WT, PH/ΔMA, Kmyr/ΔMA, and Fyn(10)/ΔMA Gag-YFP. (B) MDMs were infected with pseudotyped HIV-1 encoding WT Gag-YFP or Gag-YFP derivatives in which MA was replaced by a heterologous membrane binding sequence. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594, fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. (C) Pearson's correlation coefficients for colocalization of Gag-YFP with CD81 are shown as means and SEM. Twenty to 30 cells were analyzed per condition. n.s., not significant.
    Figure Legend Snippet: Heterologous membrane binding sequences can replace HIV-1 MA without affecting VCC localization. (A) Schematic illustrations of WT, PH/ΔMA, Kmyr/ΔMA, and Fyn(10)/ΔMA Gag-YFP. (B) MDMs were infected with pseudotyped HIV-1 encoding WT Gag-YFP or Gag-YFP derivatives in which MA was replaced by a heterologous membrane binding sequence. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594, fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. (C) Pearson's correlation coefficients for colocalization of Gag-YFP with CD81 are shown as means and SEM. Twenty to 30 cells were analyzed per condition. n.s., not significant.

    Techniques Used: Binding Assay, Infection, Sequencing, Staining, Labeling, Microscopy

    HIV-1 Gag localization to VCCs requires higher-order multimerization. (A) Schematic illustrations of WT, EE75,76AA, delNC, LZ, and LZ4 Gag-YFP. LZ4 Gag-YFP contains the WM184,185AA CA mutation, which disrupts Gag dimerization, in addition to replacement of NC with a tetramer-forming LZ sequence. (B) MDMs were infected with pseudotyped HIV-1 encoding WT Gag-YFP or Gag-YFP derivatives containing the substitutions shown in panel A. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594, fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. Note that for cells expressing delNC Gag-YFP or LZ4 Gag-YFP, prominent YFP signals were found at the cell surface as detected by ConA (white arrowheads), whereas such surface YFP signals were nearly undetectable in WT Gag-YFP-expressing cells. (C) Pearson's correlation coefficients for colocalization of Gag-YFP with ConA are shown as means and SEM. Twenty to 40 cells were analyzed per condition. *, P
    Figure Legend Snippet: HIV-1 Gag localization to VCCs requires higher-order multimerization. (A) Schematic illustrations of WT, EE75,76AA, delNC, LZ, and LZ4 Gag-YFP. LZ4 Gag-YFP contains the WM184,185AA CA mutation, which disrupts Gag dimerization, in addition to replacement of NC with a tetramer-forming LZ sequence. (B) MDMs were infected with pseudotyped HIV-1 encoding WT Gag-YFP or Gag-YFP derivatives containing the substitutions shown in panel A. At 48 h postinfection, cells were stained with ConA labeled with Alexa Fluor 594, fixed, immunostained with a mouse monoclonal anti-CD81 antibody and anti-mouse IgG conjugated with Alexa Fluor 647, and analyzed using a confocal microscope. Note that for cells expressing delNC Gag-YFP or LZ4 Gag-YFP, prominent YFP signals were found at the cell surface as detected by ConA (white arrowheads), whereas such surface YFP signals were nearly undetectable in WT Gag-YFP-expressing cells. (C) Pearson's correlation coefficients for colocalization of Gag-YFP with ConA are shown as means and SEM. Twenty to 40 cells were analyzed per condition. *, P

    Techniques Used: Mutagenesis, Sequencing, Infection, Staining, Labeling, Microscopy, Expressing

    5) Product Images from "Gag Localization and Virus-Like Particle Release Mediated by the Matrix Domain of Human T-Lymphotropic Virus Type 1 Gag Are Less Dependent on Phosphatidylinositol-(4,5)-Bisphosphate than Those Mediated by the Matrix Domain of HIV-1 Gag ▿"

    Article Title: Gag Localization and Virus-Like Particle Release Mediated by the Matrix Domain of Human T-Lymphotropic Virus Type 1 Gag Are Less Dependent on Phosphatidylinositol-(4,5)-Bisphosphate than Those Mediated by the Matrix Domain of HIV-1 Gag ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.02383-10

    Localization of both HTLV-1 and HTMA Gag remains unchanged upon 5ptaseIV overexpression, unlike that of HIV-1 Gag. (a) HeLa cells expressing Gag-eCFP constructs were fixed and stained with ConA conjugated with Alexa Fluor 594. Note that HIV-1 Gag-eCFP
    Figure Legend Snippet: Localization of both HTLV-1 and HTMA Gag remains unchanged upon 5ptaseIV overexpression, unlike that of HIV-1 Gag. (a) HeLa cells expressing Gag-eCFP constructs were fixed and stained with ConA conjugated with Alexa Fluor 594. Note that HIV-1 Gag-eCFP

    Techniques Used: Over Expression, Expressing, Construct, Staining

    6) Product Images from "Role of Nongenomic Signaling Pathways Activated by Aldosterone During Cardiac Reperfusion Injury"

    Article Title: Role of Nongenomic Signaling Pathways Activated by Aldosterone During Cardiac Reperfusion Injury

    Journal: Molecular Endocrinology

    doi: 10.1210/ME.2014-1410

    Localization of MR and GPER in LV infarcted myocardium. Representative overlay confocal microscopy images of LV sections from hearts subjected to I-R injury. MR (A) and GPR30 (B) were detected with Alexa 488 (green). Tissue sections were costained with Alexa 594 conjugate Con A (red), which specifically binds α-mannosyl saccharides in plasma membrane glycoproteins. Nuclei are stained using 4′,6′-diamino-2-phenylindole (DAPI) (blue). Scale bar, 10 μm.
    Figure Legend Snippet: Localization of MR and GPER in LV infarcted myocardium. Representative overlay confocal microscopy images of LV sections from hearts subjected to I-R injury. MR (A) and GPR30 (B) were detected with Alexa 488 (green). Tissue sections were costained with Alexa 594 conjugate Con A (red), which specifically binds α-mannosyl saccharides in plasma membrane glycoproteins. Nuclei are stained using 4′,6′-diamino-2-phenylindole (DAPI) (blue). Scale bar, 10 μm.

    Techniques Used: Confocal Microscopy, Staining

    Related Articles

    Microscopy:

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    Article Snippet: .. Only for STED microscopy on whole cells, fixed cells were labelled with 100 μg/mL concanavalin A conjugated to Alexa594 (ThermoFisher Scientific, # C11253) in HBSS (Hanks' Balanced Salt Solution; ThermoFisher Scientific, #14175053) for 30 min followed by permeabilisation for immunostaining with 0.5% Triton-X100 in PBS for 10 min prior to the blocking step. .. During immunostaining all solutions contained 0.05% Triton-X100.

    Article Title: Mitochondrial Complex V α Subunit Is Critical for Candida albicans Pathogenicity through Modulating Multiple Virulence Properties
    Article Snippet: Briefly, all strains were grown in Glass Bottom Cell Culture Dish. .. After 24 h of incubation at 37°C, the resulting biofilms were washed and stained with 25 μg/ml Concanavalin A-Alexa Fluor 594 conjugate (C-11253; Molecular Probes, Eugene, OR) at 37°C for 1 h. CSLM was performed with a Zeiss LSM 510 upright confocal microscope (Carl Zeiss, Thornwood, NY, USA), using a Zeiss Achroplan 40×, 0.8-W objective, and a HeNe1 laser with an excitation wavelength of 543 nm. .. Moreover, biofilms activity was assessed by XTT reduction assay and the crystal violet assay, accordingly to previously described protocols ( ; ; ).

    Immunostaining:

    Article Title: The packing density of a supramolecular membrane protein cluster is controlled by cytoplasmic interactions
    Article Snippet: .. Only for STED microscopy on whole cells, fixed cells were labelled with 100 μg/mL concanavalin A conjugated to Alexa594 (ThermoFisher Scientific, # C11253) in HBSS (Hanks' Balanced Salt Solution; ThermoFisher Scientific, #14175053) for 30 min followed by permeabilisation for immunostaining with 0.5% Triton-X100 in PBS for 10 min prior to the blocking step. .. During immunostaining all solutions contained 0.05% Triton-X100.

    Article Title: Efficient and gentle siRNA delivery by magnetofection
    Article Snippet: Cells were grown on coverslips in 24-well and 12-well plates, respectively, exposed to transfection procedures as outlined above and fixed with 3.7% formaldehyde. .. This was followed by a direct immunostaining protocol using concanavalin A Alexa Fluor 594 conjugate for cell surface labeling (Invitrogen). .. Nuclei were counterstained with Vectashield mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA).

    Blocking Assay:

    Article Title: The packing density of a supramolecular membrane protein cluster is controlled by cytoplasmic interactions
    Article Snippet: .. Only for STED microscopy on whole cells, fixed cells were labelled with 100 μg/mL concanavalin A conjugated to Alexa594 (ThermoFisher Scientific, # C11253) in HBSS (Hanks' Balanced Salt Solution; ThermoFisher Scientific, #14175053) for 30 min followed by permeabilisation for immunostaining with 0.5% Triton-X100 in PBS for 10 min prior to the blocking step. .. During immunostaining all solutions contained 0.05% Triton-X100.

    Article Title: O2-Filled Swimbladder Employs Monocarboxylate Transporters for the Generation of O2 by Lactate-Induced Root Effect Hemoglobin
    Article Snippet: The tissues were cryoprotected through a series of increasing sucrose concentrations up to 20% and quick frozen in optimum cutting temperature compound (Sakura Finetek, Tokyo, Japan). .. Frozen sections (6 µm) were prepared, permeabilized with 0.2% Triton X-100 in PBS at room temperature for 10 min, and incubated with 5% FBS (Invitrogen) in PBS at room temperature for 1 h. After blocking, the sections were incubated with anti-fMCT1b antiserum (1∶1,000), anti-fMCT4b antiserum (1∶5,000), and anti-GAPDH antiserum (1∶100, Sigma) in PBS containing 5% FBS at 20°C for 16 h. After washing with PBS, the sections were incubated with a mixture of Alexa Fluor 488-labeled secondary antibodies (1∶2,000), Alexa Fluor 594-labeled ConA (50 µg/ml; Invitrogen), and Hoechst 33342 (100 ng/ml) in PBS containing 5% FBS at 20°C for 1 h. The sections were mounted on antifade glycerol (90% glycerol, 10% 10× PBS, and 0.1% 1,4-phenylenediamine at pH 7.4). .. Fluorescence images were obtained as described above.

    Staining:

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    Article Title: Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector
    Article Snippet: Cells were incubated with the corresponding antibodies diluted in PBS-BSA: anti-VP2 antibody (1:200; provided by Didier Poncet, CNRS/INRA, Gif-sur-Yvette, France) directly labeled with Zenon Alexa-Fluor 594 according to the manufacturer’s instructions (Molecular Probes, Thermo Fisher Scientific), mouse monoclonal anti-VP6 (1:500; Novus Biologicals, Cambridge, UK) and the anti-mouse Alexa-Fluor 633 (1:500; Molecular Probes, Thermo Fisher Scientific), mouse monoclonal anti-V5 antibody directly labeled with fluorescein isothiocyanate (FITC) (1:500; Molecular Probes) for the V5-tagged VP7. .. The ER was stained using the Alexa-Fluor 405 conjugated lectin ConA conjugated with Alexa Fluor 594 (20 µg/µL in PBS; Molecular Probes). .. Cells were incubated with 4',6-diamidino-2-phenylindole (DAPI) (1 µg/mL in PBS, Roche, Basel, Switzerland) to visualize nuclei.

    Article Title: Mitochondrial Complex V α Subunit Is Critical for Candida albicans Pathogenicity through Modulating Multiple Virulence Properties
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    Incubation:

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    Article Snippet: The tissues were cryoprotected through a series of increasing sucrose concentrations up to 20% and quick frozen in optimum cutting temperature compound (Sakura Finetek, Tokyo, Japan). .. Frozen sections (6 µm) were prepared, permeabilized with 0.2% Triton X-100 in PBS at room temperature for 10 min, and incubated with 5% FBS (Invitrogen) in PBS at room temperature for 1 h. After blocking, the sections were incubated with anti-fMCT1b antiserum (1∶1,000), anti-fMCT4b antiserum (1∶5,000), and anti-GAPDH antiserum (1∶100, Sigma) in PBS containing 5% FBS at 20°C for 16 h. After washing with PBS, the sections were incubated with a mixture of Alexa Fluor 488-labeled secondary antibodies (1∶2,000), Alexa Fluor 594-labeled ConA (50 µg/ml; Invitrogen), and Hoechst 33342 (100 ng/ml) in PBS containing 5% FBS at 20°C for 1 h. The sections were mounted on antifade glycerol (90% glycerol, 10% 10× PBS, and 0.1% 1,4-phenylenediamine at pH 7.4). .. Fluorescence images were obtained as described above.

    Article Title: Fungistatic Action of N-Acetylcysteine on Candida albicans Biofilms and Its Interaction with Antifungal Agents
    Article Snippet: Syto9 and propidium iodide (excitation/emission at 488/488–550 nm and 488/656–700 nm, respectively) were diluted 1:1000 and incubated with the biofilm for 45 min in the dark. .. For polysaccharides (α-D-mannosyl and α-D-glucosyl) [ ], concanavalin A-Alexa Fluor 594 (Molecular Probes) (561/600–700 nm) at 75 µg/mL was incubated with the biofilms for 30 min. For β-glucans [ ], Calcofluor White (Sigma-Aldrich) (405/450 nm) at 0.5 g/L in 10% KOH was incubated with the biofilms for 10 min. For proteins [ ], biofilms were incubated with FilmTracer Sypro Ruby (Molecular Probes) (488/576–700 nm) in the dark for 30 min. For lipids [ ], Nile Red (Sigma-Aldrich) at 50 µg/mL prepared with 10 mL of 25% DMSO/H2O was incubated with the biofilm for 10 min (488/563 nm for neutral lipids and 561/613–700 nm for polar lipids). .. In order to distinguish the red fluorescence of Nile Red from the polar lipids, the green color was chosen for neutral lipids as an alternative to orange color.

    Article Title: Mitochondrial Complex V α Subunit Is Critical for Candida albicans Pathogenicity through Modulating Multiple Virulence Properties
    Article Snippet: Briefly, all strains were grown in Glass Bottom Cell Culture Dish. .. After 24 h of incubation at 37°C, the resulting biofilms were washed and stained with 25 μg/ml Concanavalin A-Alexa Fluor 594 conjugate (C-11253; Molecular Probes, Eugene, OR) at 37°C for 1 h. CSLM was performed with a Zeiss LSM 510 upright confocal microscope (Carl Zeiss, Thornwood, NY, USA), using a Zeiss Achroplan 40×, 0.8-W objective, and a HeNe1 laser with an excitation wavelength of 543 nm. .. Moreover, biofilms activity was assessed by XTT reduction assay and the crystal violet assay, accordingly to previously described protocols ( ; ; ).

    Labeling:

    Article Title: Efficient and gentle siRNA delivery by magnetofection
    Article Snippet: Cells were grown on coverslips in 24-well and 12-well plates, respectively, exposed to transfection procedures as outlined above and fixed with 3.7% formaldehyde. .. This was followed by a direct immunostaining protocol using concanavalin A Alexa Fluor 594 conjugate for cell surface labeling (Invitrogen). .. Nuclei were counterstained with Vectashield mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA).

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    Thermo Fisher concanavalin a
    CSLM images of biofilms stained with concanavalin A. Biofilms were cultured for 60 h at 37°C in Spider medium and visualized with a 40× water immersion objective, and images were reconstructed to yield views from above ( x-y  plane). (A
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    CSLM images of biofilms stained with concanavalin A. Biofilms were cultured for 60 h at 37°C in Spider medium and visualized with a 40× water immersion objective, and images were reconstructed to yield views from above ( x-y  plane). (A

    Journal:

    Article Title: Candida albicans Biofilm-Defective Mutants

    doi: 10.1128/EC.4.8.1493-1502.2005

    Figure Lengend Snippet: CSLM images of biofilms stained with concanavalin A. Biofilms were cultured for 60 h at 37°C in Spider medium and visualized with a 40× water immersion objective, and images were reconstructed to yield views from above ( x-y plane). (A

    Article Snippet: Biofilms were stained in a 2-ml solution with either 0.2 mg/ml calcofluor white (fluorescent brightener 28, F3543; Sigma) or 50 to 100 μg/ml Alexa conjugate of concanavalin A (Alexa Fluor 594 nm, C-11253; Molecular Probes ) for 1 h in the dark and observed without washing.

    Techniques: Staining, Cell Culture

    Immunohistochemistry of the fugu swimbladder. Swimbladder sections were stained with anti-MCT1b (A), anti-GAPDH (B), and anti-MCT4b (C) antisera. (D) Control samples stained with nonimmune rabbit serum. The right panels show sections double- or triple-stained with Alexa Fluor 594-labeled concanavalin A (ConA) and Hoechst 33342.

    Journal: PLoS ONE

    Article Title: O2-Filled Swimbladder Employs Monocarboxylate Transporters for the Generation of O2 by Lactate-Induced Root Effect Hemoglobin

    doi: 10.1371/journal.pone.0034579

    Figure Lengend Snippet: Immunohistochemistry of the fugu swimbladder. Swimbladder sections were stained with anti-MCT1b (A), anti-GAPDH (B), and anti-MCT4b (C) antisera. (D) Control samples stained with nonimmune rabbit serum. The right panels show sections double- or triple-stained with Alexa Fluor 594-labeled concanavalin A (ConA) and Hoechst 33342.

    Article Snippet: Frozen sections (6 µm) were prepared, permeabilized with 0.2% Triton X-100 in PBS at room temperature for 10 min, and incubated with 5% FBS (Invitrogen) in PBS at room temperature for 1 h. After blocking, the sections were incubated with anti-fMCT1b antiserum (1∶1,000), anti-fMCT4b antiserum (1∶5,000), and anti-GAPDH antiserum (1∶100, Sigma) in PBS containing 5% FBS at 20°C for 16 h. After washing with PBS, the sections were incubated with a mixture of Alexa Fluor 488-labeled secondary antibodies (1∶2,000), Alexa Fluor 594-labeled ConA (50 µg/ml; Invitrogen), and Hoechst 33342 (100 ng/ml) in PBS containing 5% FBS at 20°C for 1 h. The sections were mounted on antifade glycerol (90% glycerol, 10% 10× PBS, and 0.1% 1,4-phenylenediamine at pH 7.4).

    Techniques: Immunohistochemistry, Staining, Labeling

    Subcellular localization of MCT1b and MCT4b in the fugu swimbladder. A, High-magnification images of immunohistochemical sections of the rete mirabile stained with anti-MCT1b antiserum. The right panels show sections triple-stained with Hoechst 33342 and/or Alexa Fluor 594-labeled ConA and/or fluorescently labeled phalloidin. A, arterial capillary; V, venous capillary. B, Arterial or venous small blood vessels near the rete mirabile were stained with anti-MCT1b antiserum, Hoechst 33342, and fluorescently labeled phalloidin. A, artery; V, vein. C, High-magnification images of immunohistochemical sections of the gas gland stained with anti-MCT4b antiserum. The right panels show sections triple-stained with Alexa Fluor 594-labeled ConA and Hoechst 33342. E, endothelial cell; G, gas gland cell.

    Journal: PLoS ONE

    Article Title: O2-Filled Swimbladder Employs Monocarboxylate Transporters for the Generation of O2 by Lactate-Induced Root Effect Hemoglobin

    doi: 10.1371/journal.pone.0034579

    Figure Lengend Snippet: Subcellular localization of MCT1b and MCT4b in the fugu swimbladder. A, High-magnification images of immunohistochemical sections of the rete mirabile stained with anti-MCT1b antiserum. The right panels show sections triple-stained with Hoechst 33342 and/or Alexa Fluor 594-labeled ConA and/or fluorescently labeled phalloidin. A, arterial capillary; V, venous capillary. B, Arterial or venous small blood vessels near the rete mirabile were stained with anti-MCT1b antiserum, Hoechst 33342, and fluorescently labeled phalloidin. A, artery; V, vein. C, High-magnification images of immunohistochemical sections of the gas gland stained with anti-MCT4b antiserum. The right panels show sections triple-stained with Alexa Fluor 594-labeled ConA and Hoechst 33342. E, endothelial cell; G, gas gland cell.

    Article Snippet: Frozen sections (6 µm) were prepared, permeabilized with 0.2% Triton X-100 in PBS at room temperature for 10 min, and incubated with 5% FBS (Invitrogen) in PBS at room temperature for 1 h. After blocking, the sections were incubated with anti-fMCT1b antiserum (1∶1,000), anti-fMCT4b antiserum (1∶5,000), and anti-GAPDH antiserum (1∶100, Sigma) in PBS containing 5% FBS at 20°C for 16 h. After washing with PBS, the sections were incubated with a mixture of Alexa Fluor 488-labeled secondary antibodies (1∶2,000), Alexa Fluor 594-labeled ConA (50 µg/ml; Invitrogen), and Hoechst 33342 (100 ng/ml) in PBS containing 5% FBS at 20°C for 1 h. The sections were mounted on antifade glycerol (90% glycerol, 10% 10× PBS, and 0.1% 1,4-phenylenediamine at pH 7.4).

    Techniques: Immunohistochemistry, Staining, Labeling

    The effect of R164Q mutation on the signal transduction and cell surface expression of PKR2. A , R164Q mutation disrupted the signal transduction of PKR2 as assayed with an aequorin-based assay. B , both WT and R164Q-PKR2 were presented on the cell surface. The Alexa Fluor 594-conjugated concanavalin A was used to label the membrane. The fluorescence was monitored with a confocal microscope. Scale bar: 10 μm.

    Journal: The Journal of Biological Chemistry

    Article Title: Disease-causing Mutation in PKR2 Receptor Reveals a Critical Role of Positive Charges in the Second Intracellular Loop for G-protein Coupling and Receptor Trafficking

    doi: 10.1074/jbc.M111.223784

    Figure Lengend Snippet: The effect of R164Q mutation on the signal transduction and cell surface expression of PKR2. A , R164Q mutation disrupted the signal transduction of PKR2 as assayed with an aequorin-based assay. B , both WT and R164Q-PKR2 were presented on the cell surface. The Alexa Fluor 594-conjugated concanavalin A was used to label the membrane. The fluorescence was monitored with a confocal microscope. Scale bar: 10 μm.

    Article Snippet: Alexa Fluor 594-conjugated concanavalin A (Invitrogen) was applied to label the membrane.

    Techniques: Mutagenesis, Transduction, Expressing, Fluorescence, Microscopy

    Cellular localization of FITC-labeled siRNA (green) following magnetofection (MF; A, B) and lipofection (L2000; C, D) procedures in primary fibroblasts imaged by confocal laser scanning microscopy. Both a siRNA-only control in the absence of transfection reagent (A, 2 μg siRNA; C, 1 μg siRNA) and siRNA/transfection reagent complexes (B, D) were applied. Nuclear DNA was counterstained with DAPI (blue). The cell surface marker, ConA Alexa Fluor 594 (orange-red), was used to stain the cell membrane. The last column (“merge”) shows images from the first three columns superimposed, illustrating intracellular uptake of siRNA/transfection reagent complexes (B, D) and absence of cellular uptake of FITC-siRNA alone (A, C). Co-localization of FITC-siRNA-only or siRNA/transfection reagent complexes with the cell surface did not occur.

    Journal: Biotechnic & histochemistry : official publication of the Biological Stain Commission

    Article Title: Efficient and gentle siRNA delivery by magnetofection

    doi: 10.3109/10520291003675485

    Figure Lengend Snippet: Cellular localization of FITC-labeled siRNA (green) following magnetofection (MF; A, B) and lipofection (L2000; C, D) procedures in primary fibroblasts imaged by confocal laser scanning microscopy. Both a siRNA-only control in the absence of transfection reagent (A, 2 μg siRNA; C, 1 μg siRNA) and siRNA/transfection reagent complexes (B, D) were applied. Nuclear DNA was counterstained with DAPI (blue). The cell surface marker, ConA Alexa Fluor 594 (orange-red), was used to stain the cell membrane. The last column (“merge”) shows images from the first three columns superimposed, illustrating intracellular uptake of siRNA/transfection reagent complexes (B, D) and absence of cellular uptake of FITC-siRNA alone (A, C). Co-localization of FITC-siRNA-only or siRNA/transfection reagent complexes with the cell surface did not occur.

    Article Snippet: This was followed by a direct immunostaining protocol using concanavalin A Alexa Fluor 594 conjugate for cell surface labeling (Invitrogen).

    Techniques: Labeling, Magnetofection, Confocal Laser Scanning Microscopy, Transfection, Marker, Staining