rabbit anti-goat alexa-546 secondary antibody Search Results


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  • 99
    Thermo Fisher goat anti rabbit igg h l cross adsorbed secondary antibody
    Outcomes of modulating functional IMPα2/α4/α6 levels on paraspeckle marker (endogenous PSPC1/SFPQ or exogenous DsRed2-PSPC1) nuclear transport and paraspeckle localization in HeLa cells. HeLa cells were transiently transfected with constructs encoding GFP-tagged IMPα2/α4/α6 variants (see Fig. 1A for predicted function) as indicated, with labels at the bottom of each panel and consistent colors used throughout. Paraspeckles were assessed within experimental groups using indirect immunofluorescence with an <t>Alexa</t> <t>Fluor</t> 546 (A546) secondary antibody to detect endogenous PSPC1 ( A ), using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous SFPQ ( B ) or through exogenous PSPC1 by co-transfecting with a plasmid encoding DsRed2-PSPC1 ( C ). After analysis pipeline as outlined in Fig. 1B , the primary measures are presented as bar graphs and scatter plots with overlaid box plots indicating the mean and interquartile ranges for each experimental group. Samples with statistically significant differences within IMPα groups are indicated (*). The primary measures include the percentage of foci positive cells (i), the ratio fluorescent signal within the nucleus and cytoplasm (F n/c ) for the paraspeckle marker used (ii), the number of foci detected per cell (iii), the sum foci associated fluorescent signal per cell for the paraspeckle marker used (iv) and the sum foci associated fluorescent signal per foci for the paraspeckle marker used (v). These measures were assessed within groups per entire cell population (i, ii), per foci positive cell populations (iii, iv) or on a per foci basis (v).
    Goat Anti Rabbit Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14437 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti rabbit igg h l highly cross adsorbed secondary antibody
    Outcomes of modulating functional IMPα2/α4/α6 levels on paraspeckle marker (endogenous PSPC1/SFPQ or exogenous DsRed2-PSPC1) nuclear transport and paraspeckle localization in HeLa cells. HeLa cells were transiently transfected with constructs encoding GFP-tagged IMPα2/α4/α6 variants (see Fig. 1A for predicted function) as indicated, with labels at the bottom of each panel and consistent colors used throughout. Paraspeckles were assessed within experimental groups using indirect immunofluorescence with an <t>Alexa</t> <t>Fluor</t> 546 (A546) secondary antibody to detect endogenous PSPC1 ( A ), using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous SFPQ ( B ) or through exogenous PSPC1 by co-transfecting with a plasmid encoding DsRed2-PSPC1 ( C ). After analysis pipeline as outlined in Fig. 1B , the primary measures are presented as bar graphs and scatter plots with overlaid box plots indicating the mean and interquartile ranges for each experimental group. Samples with statistically significant differences within IMPα groups are indicated (*). The primary measures include the percentage of foci positive cells (i), the ratio fluorescent signal within the nucleus and cytoplasm (F n/c ) for the paraspeckle marker used (ii), the number of foci detected per cell (iii), the sum foci associated fluorescent signal per cell for the paraspeckle marker used (iv) and the sum foci associated fluorescent signal per foci for the paraspeckle marker used (v). These measures were assessed within groups per entire cell population (i, ii), per foci positive cell populations (iii, iv) or on a per foci basis (v).
    Goat Anti Rabbit Igg H L Highly Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 10173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher alexa 546 conjugated goat anti rabbit secondary antibody
    IDO1 protein measurement in hormone receptor-positive breast cancer. Fluorescent microphotographs of representative hormone receptor-positive breast cancer cases showing different levels of IDO1 protein. Staining pattern was cytoplasmic. Blue: 4,6-diamidino-2-phenylindole (DAPI). Green: <t>Alexa-546</t> (Cytokeratin [CK]). Red: Cyanine-5 (IDO1). Scale bars: 100 um
    Alexa 546 Conjugated Goat Anti Rabbit Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 121 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 546 goat anti rabbit secondary antibody
    IDO1 protein measurement in hormone receptor-positive breast cancer. Fluorescent microphotographs of representative hormone receptor-positive breast cancer cases showing different levels of IDO1 protein. Staining pattern was cytoplasmic. Blue: 4,6-diamidino-2-phenylindole (DAPI). Green: <t>Alexa-546</t> (Cytokeratin [CK]). Red: Cyanine-5 (IDO1). Scale bars: 100 um
    Alexa Fluor 546 Goat Anti Rabbit Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher secondary antibody
    IDO1 protein measurement in hormone receptor-positive breast cancer. Fluorescent microphotographs of representative hormone receptor-positive breast cancer cases showing different levels of IDO1 protein. Staining pattern was cytoplasmic. Blue: 4,6-diamidino-2-phenylindole (DAPI). Green: <t>Alexa-546</t> (Cytokeratin [CK]). Red: Cyanine-5 (IDO1). Scale bars: 100 um
    Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher f ab 2 goat anti rabbit igg h l cross adsorbed secondary antibody
    IDO1 protein measurement in hormone receptor-positive breast cancer. Fluorescent microphotographs of representative hormone receptor-positive breast cancer cases showing different levels of IDO1 protein. Staining pattern was cytoplasmic. Blue: 4,6-diamidino-2-phenylindole (DAPI). Green: <t>Alexa-546</t> (Cytokeratin [CK]). Red: Cyanine-5 (IDO1). Scale bars: 100 um
    F Ab 2 Goat Anti Rabbit Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1504 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 546 conjugated goat anti rabbit secondary antibody
    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and <t>Alexa</t> Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.
    Alexa Fluor 546 Conjugated Goat Anti Rabbit Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 546
    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and <t>Alexa</t> Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.
    Alexa Fluor 546, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5720 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 546 conjugated goat anti rabbit igg secondary antibody
    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and <t>Alexa</t> Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.
    Alexa Fluor 546 Conjugated Goat Anti Rabbit Igg Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Jackson Immuno alexa fluor 488 affinipure f ab 2 fragment goat anti rabbit igg
    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and <t>Alexa</t> Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.
    Alexa Fluor 488 Affinipure F Ab 2 Fragment Goat Anti Rabbit Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 94/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti rat igg h l cross adsorbed secondary antibody
    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and <t>Alexa</t> Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.
    Goat Anti Rat Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3571 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rabbit anti goat igg h l cross adsorbed secondary antibody
    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and <t>Alexa</t> Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.
    Rabbit Anti Goat Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 453 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Outcomes of modulating functional IMPα2/α4/α6 levels on paraspeckle marker (endogenous PSPC1/SFPQ or exogenous DsRed2-PSPC1) nuclear transport and paraspeckle localization in HeLa cells. HeLa cells were transiently transfected with constructs encoding GFP-tagged IMPα2/α4/α6 variants (see Fig. 1A for predicted function) as indicated, with labels at the bottom of each panel and consistent colors used throughout. Paraspeckles were assessed within experimental groups using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous PSPC1 ( A ), using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous SFPQ ( B ) or through exogenous PSPC1 by co-transfecting with a plasmid encoding DsRed2-PSPC1 ( C ). After analysis pipeline as outlined in Fig. 1B , the primary measures are presented as bar graphs and scatter plots with overlaid box plots indicating the mean and interquartile ranges for each experimental group. Samples with statistically significant differences within IMPα groups are indicated (*). The primary measures include the percentage of foci positive cells (i), the ratio fluorescent signal within the nucleus and cytoplasm (F n/c ) for the paraspeckle marker used (ii), the number of foci detected per cell (iii), the sum foci associated fluorescent signal per cell for the paraspeckle marker used (iv) and the sum foci associated fluorescent signal per foci for the paraspeckle marker used (v). These measures were assessed within groups per entire cell population (i, ii), per foci positive cell populations (iii, iv) or on a per foci basis (v).

    Journal: Scientific Reports

    Article Title: Development of a pipeline for automated, high-throughput analysis of paraspeckle proteins reveals specific roles for importin α proteins

    doi: 10.1038/srep43323

    Figure Lengend Snippet: Outcomes of modulating functional IMPα2/α4/α6 levels on paraspeckle marker (endogenous PSPC1/SFPQ or exogenous DsRed2-PSPC1) nuclear transport and paraspeckle localization in HeLa cells. HeLa cells were transiently transfected with constructs encoding GFP-tagged IMPα2/α4/α6 variants (see Fig. 1A for predicted function) as indicated, with labels at the bottom of each panel and consistent colors used throughout. Paraspeckles were assessed within experimental groups using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous PSPC1 ( A ), using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous SFPQ ( B ) or through exogenous PSPC1 by co-transfecting with a plasmid encoding DsRed2-PSPC1 ( C ). After analysis pipeline as outlined in Fig. 1B , the primary measures are presented as bar graphs and scatter plots with overlaid box plots indicating the mean and interquartile ranges for each experimental group. Samples with statistically significant differences within IMPα groups are indicated (*). The primary measures include the percentage of foci positive cells (i), the ratio fluorescent signal within the nucleus and cytoplasm (F n/c ) for the paraspeckle marker used (ii), the number of foci detected per cell (iii), the sum foci associated fluorescent signal per cell for the paraspeckle marker used (iv) and the sum foci associated fluorescent signal per foci for the paraspeckle marker used (v). These measures were assessed within groups per entire cell population (i, ii), per foci positive cell populations (iii, iv) or on a per foci basis (v).

    Article Snippet: Secondary antibodies, rabbit anti-mouse Alexa Fluor 546 (Molecular Probes-Invitrogen, cat#A11060) for GFP/RFP-tagged transfections and donkey anti-mouse Alexa Fluor 488 (Molecular Probes-Invitrogen, cat#A21202) plus goat anti-rabbit Alexa Fluor 546 (Molecular Probes-Invitrogen, cat#A11010) or rabbit anti-goat Alexa Fluor 546 (Molecular Probes-Invitrogen, cat#A21085) for siRNA knockdown samples (1:200 in 0.5% BSA/PBS), were applied for 90 mins at room temperature.

    Techniques: Functional Assay, Marker, Transfection, Construct, Immunofluorescence, Plasmid Preparation

    Modulating functional IMPα2/α4/α6 levels affected a plethora measurable paraspeckle related outcomes that can be simultaneously visualised using principal component analysis (PCA). The results of transiently transfecting HeLa cells with constructs encoding GFP-tagged IMPα2/α4/α6 variants ( Tables 1 , 2 and 3 and Fig. 2 ) were used to perform PCA, allowing simultaneous comparisons of multiple parameters and revealing strong patterns between groups. In each experiment PC1 explains > 99% of the variance across all parameter and therefore the distances between groups across the X axis (PC1) should be considered as the primary delineator. Paraspeckles were assessed within experimental groups using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous PSPC1 ( A ), using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous SFPQ ( B ) or through exogenous PSPC1 by co-transfecting with a plasmid encoding DsRed2-PSPC1 ( C ). Parameters used to compare the geometric means of groups within experiments using a specific paraspeckle marker (PSM; A:PSPC1, B:SFPQ, C:DsRed2-PSPC1) were “% cells positive for foci”, “cytoplasmic PSM intensity”, “nuclear PSM intensity”, “PSM F n/c per cell”, “PSM intensity per cell”, “number of nuclear foci per cell”, “sum volume of nuclear foci per cell”, “sum nuclear foci PSM intensity per cell”, “nuclear foci volume”, “nuclear foci PSM intensity” and “sum nuclear foci PSM intensity”.

    Journal: Scientific Reports

    Article Title: Development of a pipeline for automated, high-throughput analysis of paraspeckle proteins reveals specific roles for importin α proteins

    doi: 10.1038/srep43323

    Figure Lengend Snippet: Modulating functional IMPα2/α4/α6 levels affected a plethora measurable paraspeckle related outcomes that can be simultaneously visualised using principal component analysis (PCA). The results of transiently transfecting HeLa cells with constructs encoding GFP-tagged IMPα2/α4/α6 variants ( Tables 1 , 2 and 3 and Fig. 2 ) were used to perform PCA, allowing simultaneous comparisons of multiple parameters and revealing strong patterns between groups. In each experiment PC1 explains > 99% of the variance across all parameter and therefore the distances between groups across the X axis (PC1) should be considered as the primary delineator. Paraspeckles were assessed within experimental groups using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous PSPC1 ( A ), using indirect immunofluorescence with an Alexa Fluor 546 (A546) secondary antibody to detect endogenous SFPQ ( B ) or through exogenous PSPC1 by co-transfecting with a plasmid encoding DsRed2-PSPC1 ( C ). Parameters used to compare the geometric means of groups within experiments using a specific paraspeckle marker (PSM; A:PSPC1, B:SFPQ, C:DsRed2-PSPC1) were “% cells positive for foci”, “cytoplasmic PSM intensity”, “nuclear PSM intensity”, “PSM F n/c per cell”, “PSM intensity per cell”, “number of nuclear foci per cell”, “sum volume of nuclear foci per cell”, “sum nuclear foci PSM intensity per cell”, “nuclear foci volume”, “nuclear foci PSM intensity” and “sum nuclear foci PSM intensity”.

    Article Snippet: Secondary antibodies, rabbit anti-mouse Alexa Fluor 546 (Molecular Probes-Invitrogen, cat#A11060) for GFP/RFP-tagged transfections and donkey anti-mouse Alexa Fluor 488 (Molecular Probes-Invitrogen, cat#A21202) plus goat anti-rabbit Alexa Fluor 546 (Molecular Probes-Invitrogen, cat#A11010) or rabbit anti-goat Alexa Fluor 546 (Molecular Probes-Invitrogen, cat#A21085) for siRNA knockdown samples (1:200 in 0.5% BSA/PBS), were applied for 90 mins at room temperature.

    Techniques: Functional Assay, Construct, Immunofluorescence, Plasmid Preparation, Marker

    Loss of VCP induces EMT A. 72 hours after transfection of A549, H358 or immortalized human bronchiolar HPLD-1 cells with siRNA against VCP or non-targeting cell lysates were prepared and western blot analysis of EMT markers Vimentin and E-cadherin was performed. B. Fluorescence staining for E-cadherin and Vimentin in A549 cells. After 24 hrs of transfection either with non-targeting siRNA (siNT) or with siRNAs targeting VCP (siVCP1 or siVCP2) cells were trypsinized and plated on chamber slides and stained for EMT markers. i, iii and v: E-cadherin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). ii, iv and vi: overlay of respective E-cadherin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a, c and e: Vimentin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b, d and f: overlay of respective Vimentin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. C. A549 cells were prepared as described in B and F-actin was detected with Alexa Fluor 568 Phalloidin (red). Re-organization of actin cytoskeleton through destruction and cellular protrusion formation is indicated by arrows.

    Journal: Oncotarget

    Article Title: Regulation of VCP/p97 demonstrates the critical balance between cell death and epithelial-mesenchymal transition (EMT) downstream of ER stress

    doi:

    Figure Lengend Snippet: Loss of VCP induces EMT A. 72 hours after transfection of A549, H358 or immortalized human bronchiolar HPLD-1 cells with siRNA against VCP or non-targeting cell lysates were prepared and western blot analysis of EMT markers Vimentin and E-cadherin was performed. B. Fluorescence staining for E-cadherin and Vimentin in A549 cells. After 24 hrs of transfection either with non-targeting siRNA (siNT) or with siRNAs targeting VCP (siVCP1 or siVCP2) cells were trypsinized and plated on chamber slides and stained for EMT markers. i, iii and v: E-cadherin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). ii, iv and vi: overlay of respective E-cadherin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a, c and e: Vimentin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b, d and f: overlay of respective Vimentin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. C. A549 cells were prepared as described in B and F-actin was detected with Alexa Fluor 568 Phalloidin (red). Re-organization of actin cytoskeleton through destruction and cellular protrusion formation is indicated by arrows.

    Article Snippet: USA) or Alexa Fluor 488 goat anti-rabbit IgG #A11034 or Alexa Fluor 546 goat anti-rabbit IgG #A11010 (Molecular Probes, Invitrogen detection technologies, Eugene, OR.

    Techniques: Transfection, Western Blot, Fluorescence, Staining

    Loss of VCP induces ER stress A. Western blot analysis of different proteins involved in unfolded protein response and ER stress response. Cells were transfected with either non targeting (siNT) siRNA or two different siRNAs targeting VCP (siVCP1 and siVCP2). After 72 hrs of transfection, cells were harvested and analyzed for protein expression. B. Fluorescence staining for Chop and Calnexin in A549 and H358 cells. After 24 hrs of transfection either with non-targeting siRNA (siNT) or with siRNAs targeting VCP (siVCP1 or siVCP2) cells were trypsinized and plated on chamber slides. After 48 hours cells were fixed and stained for Chop and Calnexin. i, iii and v: Chop was detected using Alexa Fluor 488 rabbit anti-mouse IgG (green). ii, iv and vi: overlay of respective Chop and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a, c and e: Calnexin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b, d and f: overlay of respective Calnexin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain.

    Journal: Oncotarget

    Article Title: Regulation of VCP/p97 demonstrates the critical balance between cell death and epithelial-mesenchymal transition (EMT) downstream of ER stress

    doi:

    Figure Lengend Snippet: Loss of VCP induces ER stress A. Western blot analysis of different proteins involved in unfolded protein response and ER stress response. Cells were transfected with either non targeting (siNT) siRNA or two different siRNAs targeting VCP (siVCP1 and siVCP2). After 72 hrs of transfection, cells were harvested and analyzed for protein expression. B. Fluorescence staining for Chop and Calnexin in A549 and H358 cells. After 24 hrs of transfection either with non-targeting siRNA (siNT) or with siRNAs targeting VCP (siVCP1 or siVCP2) cells were trypsinized and plated on chamber slides. After 48 hours cells were fixed and stained for Chop and Calnexin. i, iii and v: Chop was detected using Alexa Fluor 488 rabbit anti-mouse IgG (green). ii, iv and vi: overlay of respective Chop and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain. a, c and e: Calnexin was detected using Alexa Fluor 488 goat anti-rabbit IgG (green). b, d and f: overlay of respective Calnexin and F-actin (Alexa Fluor 568 Phalloidin; red) staining with DAPI counter stain.

    Article Snippet: USA) or Alexa Fluor 488 goat anti-rabbit IgG #A11034 or Alexa Fluor 546 goat anti-rabbit IgG #A11010 (Molecular Probes, Invitrogen detection technologies, Eugene, OR.

    Techniques: Western Blot, Transfection, Expressing, Fluorescence, Staining

    MK-0524 induces translocation of DP1 from intracellular compartments to the plasma membrane. The distribution of DP1 in HEK293 cells was determined by immunofluorescence confocal microscopy. HEK293 cells transfected with Flag-DP1 were treated with vehicle or 1 µM of MK-0524 alone or in presence of 20 µM Brefeldin A for 90 min. Cells were labeled with mouse anti-FLAG and either a rabbit anti-calnexin antibody (top and midlle panels) or a rabbit anti-protein disulfide isomerase (PDI) antibody (lower panel) as described under “ Materials and Methods ”. Secondary antibodies were Alexa Fluor 488 donkey anti-mouse IgG and Alexa Fluor 546 goat anti-rabbit IgG. Merge images of the green-labelled DP1 and red-labeled calnexin or PDI are shown. Bars, 10 µM.

    Journal: PLoS ONE

    Article Title: Inverse Agonist and Pharmacochaperone Properties of MK-0524 on the Prostanoid DP1 Receptor

    doi: 10.1371/journal.pone.0065767

    Figure Lengend Snippet: MK-0524 induces translocation of DP1 from intracellular compartments to the plasma membrane. The distribution of DP1 in HEK293 cells was determined by immunofluorescence confocal microscopy. HEK293 cells transfected with Flag-DP1 were treated with vehicle or 1 µM of MK-0524 alone or in presence of 20 µM Brefeldin A for 90 min. Cells were labeled with mouse anti-FLAG and either a rabbit anti-calnexin antibody (top and midlle panels) or a rabbit anti-protein disulfide isomerase (PDI) antibody (lower panel) as described under “ Materials and Methods ”. Secondary antibodies were Alexa Fluor 488 donkey anti-mouse IgG and Alexa Fluor 546 goat anti-rabbit IgG. Merge images of the green-labelled DP1 and red-labeled calnexin or PDI are shown. Bars, 10 µM.

    Article Snippet: Alexa Fluor 488 donkey anti-mouse (cat. A-21202) and Alexa Fluor 546 goat anti-rabbit (cat. A-11035) secondary antibodies were from Molecular Probes, CA.

    Techniques: Translocation Assay, Immunofluorescence, Confocal Microscopy, Transfection, Labeling

    CALX is highly expressed in the outer dendrites of olfactory sensory neurons (OSNs). Superior panels:  (A,D)  GCaMP6f staining with Alexa 488.  (B,E)  CALX staining with Alexa 546.  (C,F)  Merged stainings.  (A–C)  General overview of female  Drosophila  antennae. Bars, 20 μm.  (D–F)  Details of a section at higher magnification, Bars, 5 μm. White arrows indicate localization of CALX in the dendrites. Lower panels:  (G,J)  GCaMP6f staining with Alexa 488.  (H,K)  Control staining with only Alexa 546.  (I,L)  Merged stainings.  (G–I)  General overview of female  Drosophila  antennae. Bars, 20 μm.  (J–L)  Details of a section at higher magnification, Bars, 5 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: The Drosophila melanogaster Na+/Ca2+ Exchanger CALX Controls the Ca2+ Level in Olfactory Sensory Neurons at Rest and After Odorant Receptor Activation

    doi: 10.3389/fncel.2018.00186

    Figure Lengend Snippet: CALX is highly expressed in the outer dendrites of olfactory sensory neurons (OSNs). Superior panels: (A,D) GCaMP6f staining with Alexa 488. (B,E) CALX staining with Alexa 546. (C,F) Merged stainings. (A–C) General overview of female Drosophila antennae. Bars, 20 μm. (D–F) Details of a section at higher magnification, Bars, 5 μm. White arrows indicate localization of CALX in the dendrites. Lower panels: (G,J) GCaMP6f staining with Alexa 488. (H,K) Control staining with only Alexa 546. (I,L) Merged stainings. (G–I) General overview of female Drosophila antennae. Bars, 20 μm. (J–L) Details of a section at higher magnification, Bars, 5 μm.

    Article Snippet: Secondary antibodies used: goat anti mouse Alexa 488- and goat anti rabbit Alexa 546 (A-11120, Invitrogen, Carlsbad, CA, USA).

    Techniques: Staining

    Starvation augments the colocalization of GFP-LC3 with TRPV1. (A–L) HeLa cells were transiently transfected with pGFP-LC3. After 2 days, cells were incubated in either the absence or presence of CLQ for 6 h in normal medium containing serum (A–C and G–I) or in HBBS to stimulate starvation (D–F and J–L). Control represents DMSO treatment instead of CLQ. Immunofluorescence was performed using rabbit anti-TRPV1 and Alexa Fluor 546 goat anti-rabbit IgG antibody for TRPV1 (red), GFP-LC3 (green) and DAPI for nuclei (blue). White arrow heads indicate the regions of colocalization of GFP-LC3 with TRPV1. (Right panel) Colocalization of GFPLC3 with TRPV1 in the left panel (C, F, I, and L) was quantified. Columns and error bars represent the mean values and standard deviations. The numbers above the error bars indicate the mean values. ** P

    Journal: Molecules and Cells

    Article Title: Transient Receptor Potential Cation Channel V1 (TRPV1) Is Degraded by Starvation- and Glucocorticoid-Mediated Autophagy

    doi: 10.14348/molcells.2014.2384

    Figure Lengend Snippet: Starvation augments the colocalization of GFP-LC3 with TRPV1. (A–L) HeLa cells were transiently transfected with pGFP-LC3. After 2 days, cells were incubated in either the absence or presence of CLQ for 6 h in normal medium containing serum (A–C and G–I) or in HBBS to stimulate starvation (D–F and J–L). Control represents DMSO treatment instead of CLQ. Immunofluorescence was performed using rabbit anti-TRPV1 and Alexa Fluor 546 goat anti-rabbit IgG antibody for TRPV1 (red), GFP-LC3 (green) and DAPI for nuclei (blue). White arrow heads indicate the regions of colocalization of GFP-LC3 with TRPV1. (Right panel) Colocalization of GFPLC3 with TRPV1 in the left panel (C, F, I, and L) was quantified. Columns and error bars represent the mean values and standard deviations. The numbers above the error bars indicate the mean values. ** P

    Article Snippet: Next, cells were incubated with Alexa Fluor 546 goat anti-rabbit IgG secondary antibodies (Invitrogen).

    Techniques: Transfection, Incubation, Immunofluorescence

    IDO1 protein measurement in hormone receptor-positive breast cancer. Fluorescent microphotographs of representative hormone receptor-positive breast cancer cases showing different levels of IDO1 protein. Staining pattern was cytoplasmic. Blue: 4,6-diamidino-2-phenylindole (DAPI). Green: Alexa-546 (Cytokeratin [CK]). Red: Cyanine-5 (IDO1). Scale bars: 100 um

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Objective measurement and clinical significance of IDO1 protein in hormone receptor-positive breast cancer

    doi: 10.1186/s40425-017-0285-7

    Figure Lengend Snippet: IDO1 protein measurement in hormone receptor-positive breast cancer. Fluorescent microphotographs of representative hormone receptor-positive breast cancer cases showing different levels of IDO1 protein. Staining pattern was cytoplasmic. Blue: 4,6-diamidino-2-phenylindole (DAPI). Green: Alexa-546 (Cytokeratin [CK]). Red: Cyanine-5 (IDO1). Scale bars: 100 um

    Article Snippet: Sections were then incubated for one hour at room temperature with Alexa 546-conjugated goat anti-rabbit secondary antibody (Molecular Probes, Eugene, OR, USA) diluted 1:100 in mouse EnVision amplification reagent (Dako), or Alexa 546-conjugated goat anti-mouse secondary antibody (Molecular Probes) in 1:100 rabbit Envision amplification reagent (Dako).

    Techniques: Staining

    PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and Alexa Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.

    Journal: Journal of Biochemistry

    Article Title: PRMT2 interacts with splicing factors and regulates the alternative splicing of BCL-X

    doi: 10.1093/jb/mvw102

    Figure Lengend Snippet: PRMT2 affects the subcellular localization of SAM68. HeLa cells untransfected ( A ) or transfected with mCitrine-PRMT2 ( B , green ) or mCitrine-ΔSH3 ( C , green ) were treated with or without 20 μM AdOx as indicated. SAM68 was immunostained with anti-SAM68 and Alexa Fluor 546-conjugated antibodies ( red ). Nuclei were visualized by DAPI stain ( blue ). The phase contrasted images ( left ) and the merged images ( right ) are also shown. The scale bar indicates 10 µm.

    Article Snippet: The cells were first blotted with an anti-SAM68 antibody at 1:50 dilution, then with an Alexa Fluor 546-conjugated goat anti-rabbit secondary antibody (Invitrogen) at a 1:1,000 dilution, both in 1% BSA (w/v) in PBS with 0.1% Tween-20 (v/v) (PBST) at room temperature for 1 h with three 5-min washes between each step.

    Techniques: Transfection, Staining