goat anti rabbit igg alexa fluor 568 conjugate  (Thermo Fisher)


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    Structured Review

    Thermo Fisher goat anti rabbit igg alexa fluor 568 conjugate
    Validation of UQCRC1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human UQCRC1, followed by goat anti-rabbit <t>IgG–Alexa</t> Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).
    Goat Anti Rabbit Igg Alexa Fluor 568 Conjugate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg alexa fluor 568 conjugate/product/Thermo Fisher
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg alexa fluor 568 conjugate - by Bioz Stars, 2020-07
    90/100 stars

    Images

    1) Product Images from "Identification of specific biomarkers for gastric adenocarcinoma by ITRAQ proteomic approach"

    Article Title: Identification of specific biomarkers for gastric adenocarcinoma by ITRAQ proteomic approach

    Journal: Scientific Reports

    doi: 10.1038/srep38871

    Validation of UQCRC1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human UQCRC1, followed by goat anti-rabbit IgG–Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).
    Figure Legend Snippet: Validation of UQCRC1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human UQCRC1, followed by goat anti-rabbit IgG–Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).

    Techniques Used: Expressing, Immunofluorescence, Staining, Incubation

    Validation of Annexin A1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human Annexin A1, followed by goat anti-rabbit IgG - Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).
    Figure Legend Snippet: Validation of Annexin A1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human Annexin A1, followed by goat anti-rabbit IgG - Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).

    Techniques Used: Expressing, Immunofluorescence, Staining, Incubation

    2) Product Images from "Novel MAG Variant Causes Cerebellar Ataxia with Oculomotor Apraxia: Molecular Basis and Expanded Clinical Phenotype"

    Article Title: Novel MAG Variant Causes Cerebellar Ataxia with Oculomotor Apraxia: Molecular Basis and Expanded Clinical Phenotype

    Journal: Journal of Clinical Medicine

    doi: 10.3390/jcm9041212

    MAG-C42R has an altered subcellular localization. ( A ) The localization of EGFP-tagged MAG clones in HEK293T cells. EGFP was directly visualized. The presence of MAG in the endoplasmic reticulum and Golgi complex was analyzed by probing cells with anti-calnexin and anti-GM130 antibodies, respectively, both detected with Alexa Fluor 568-conjugated secondary antibodies (red). DNA was stained with Hoechst 33342 (blue). Photographs were acquired using a Zeiss Axio Imager Z1 microscope. Bars: 10 μm. ( B ) The co-immunoprecipitation of EGFP-tagged MAG clones with endogenous calnexin and GM130 in HEK293T cells. Cell lysates were immunoprecipitated with anti-GFP antibody. The immunoprecipitates were subjected to immunoblotting with anti-GM130 antibody, then stripped and reprobed with anti-calnexin antibody, followed by stripping and reprobing with anti-EGFP antibody. The same methodology was applied to whole cell lysates. Calnexin and GM130 immunoblotting figures have different exposure times (original blots are presented in  Figure S2 ).
    Figure Legend Snippet: MAG-C42R has an altered subcellular localization. ( A ) The localization of EGFP-tagged MAG clones in HEK293T cells. EGFP was directly visualized. The presence of MAG in the endoplasmic reticulum and Golgi complex was analyzed by probing cells with anti-calnexin and anti-GM130 antibodies, respectively, both detected with Alexa Fluor 568-conjugated secondary antibodies (red). DNA was stained with Hoechst 33342 (blue). Photographs were acquired using a Zeiss Axio Imager Z1 microscope. Bars: 10 μm. ( B ) The co-immunoprecipitation of EGFP-tagged MAG clones with endogenous calnexin and GM130 in HEK293T cells. Cell lysates were immunoprecipitated with anti-GFP antibody. The immunoprecipitates were subjected to immunoblotting with anti-GM130 antibody, then stripped and reprobed with anti-calnexin antibody, followed by stripping and reprobing with anti-EGFP antibody. The same methodology was applied to whole cell lysates. Calnexin and GM130 immunoblotting figures have different exposure times (original blots are presented in Figure S2 ).

    Techniques Used: Clone Assay, Staining, Microscopy, Immunoprecipitation, Stripping Membranes

    Related Articles

    Staining:

    Article Title: Identification of specific biomarkers for gastric adenocarcinoma by ITRAQ proteomic approach
    Article Snippet: .. After washing, the sections were incubated with goat anti-rabbit IgG - Alexa Fluor® 568 conjugate (Thermo Fisher Scientific), or goat anti-mouse IgG- Alexa Fluor® 488 conjugate, followed by incubation with DAPI to stain nuclei. .. Images were acquired using a Multiphoton Laser Scanning Microscope (FV1000, Olympus).

    Article Title: Downregulation of HADH promotes gastric cancer progression via Akt signaling pathway
    Article Snippet: .. After washing, the sections were incubated with goat anti-rabbit IgG - Alexa Fluor® 568 conjugate (Thermo Fisher Scientific), followed by incubation with DAPI to stain nuclei. .. Images were acquired using a Multiphoton Laser Scanning Microscope (FV1000, Olympus).

    Incubation:

    Article Title: Identification of specific biomarkers for gastric adenocarcinoma by ITRAQ proteomic approach
    Article Snippet: .. After washing, the sections were incubated with goat anti-rabbit IgG - Alexa Fluor® 568 conjugate (Thermo Fisher Scientific), or goat anti-mouse IgG- Alexa Fluor® 488 conjugate, followed by incubation with DAPI to stain nuclei. .. Images were acquired using a Multiphoton Laser Scanning Microscope (FV1000, Olympus).

    Article Title: Downregulation of HADH promotes gastric cancer progression via Akt signaling pathway
    Article Snippet: .. After washing, the sections were incubated with goat anti-rabbit IgG - Alexa Fluor® 568 conjugate (Thermo Fisher Scientific), followed by incubation with DAPI to stain nuclei. .. Images were acquired using a Multiphoton Laser Scanning Microscope (FV1000, Olympus).

    other:

    Article Title: Novel MAG Variant Causes Cerebellar Ataxia with Oculomotor Apraxia: Molecular Basis and Expanded Clinical Phenotype
    Article Snippet: Secondary antibodies: horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (sc-2005, Santa Cruz Biotechnology, Heidelberg, Germany), HRP-conjugated goat anti-rabbit IgG (401393, Calbiochem, EDM Millipore, Darmstadt, Germany,), goat anti-mouse IgG Alexa Fluor® 568 conjugate (A-11004, ThermoFisher Scientific, Waltham, MA, USA), and goat anti-rabbit IgG Alexa Fluor® 568 conjugate (A-11011, ThermoFisher Scientific, Waltham, MA, USA).

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    Thermo Fisher goat anti rabbit igg alexa fluor 568 conjugate
    Validation of UQCRC1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human UQCRC1, followed by goat anti-rabbit <t>IgG–Alexa</t> Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).
    Goat Anti Rabbit Igg Alexa Fluor 568 Conjugate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg alexa fluor 568 conjugate/product/Thermo Fisher
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg alexa fluor 568 conjugate - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    84
    Thermo Fisher goat anti rabbit igg alexafluor 568 conjugated secondary antibody
    Twinfilin regulates capping protein localization and dynamics. (A) Localization of EGFP-CP in wild-type and twf1/twf2-KO B16-F1 cells, where F-actin was visualized with <t>AlexaFluor-568</t> phalloidin. Panels in the middle and right are magnifications of lamellipodial regions highlighted in the whole cell images in left. Scale bars = 10 μ m. (B) Examples of line profiles generated across the center of lamellipodia as indicated with dotted lines. Data represent mean of 5 measurements of individual lamellipodia, with standard deviations shown. The ‘0 μ m’ value in x-axis is set to correspond the peak intensity of phalloidin. (C) The ratio of CP and F-actin co-localization widths were detected by measuring the width of localization at 50% of maximum intensity. Data points represent measurements from individual lamellipodia with mean values and standard deviations shown. Statistical significance was calculated with Student’s unpaired, two-tailed t-test. ****, p
    Goat Anti Rabbit Igg Alexafluor 568 Conjugated Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg alexafluor 568 conjugated secondary antibody/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg alexafluor 568 conjugated secondary antibody - by Bioz Stars, 2020-07
    84/100 stars
      Buy from Supplier

    91
    Thermo Fisher alexa fluor 568 conjugated goat anti rabbit igg
    Twinfilin regulates capping protein localization and dynamics. (A) Localization of EGFP-CP in wild-type and twf1/twf2-KO B16-F1 cells, where F-actin was visualized with <t>AlexaFluor-568</t> phalloidin. Panels in the middle and right are magnifications of lamellipodial regions highlighted in the whole cell images in left. Scale bars = 10 μ m. (B) Examples of line profiles generated across the center of lamellipodia as indicated with dotted lines. Data represent mean of 5 measurements of individual lamellipodia, with standard deviations shown. The ‘0 μ m’ value in x-axis is set to correspond the peak intensity of phalloidin. (C) The ratio of CP and F-actin co-localization widths were detected by measuring the width of localization at 50% of maximum intensity. Data points represent measurements from individual lamellipodia with mean values and standard deviations shown. Statistical significance was calculated with Student’s unpaired, two-tailed t-test. ****, p
    Alexa Fluor 568 Conjugated Goat Anti Rabbit Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa fluor 568 conjugated goat anti rabbit igg/product/Thermo Fisher
    Average 91 stars, based on 114 article reviews
    Price from $9.99 to $1999.99
    alexa fluor 568 conjugated goat anti rabbit igg - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    Image Search Results


    Validation of UQCRC1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human UQCRC1, followed by goat anti-rabbit IgG–Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).

    Journal: Scientific Reports

    Article Title: Identification of specific biomarkers for gastric adenocarcinoma by ITRAQ proteomic approach

    doi: 10.1038/srep38871

    Figure Lengend Snippet: Validation of UQCRC1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human UQCRC1, followed by goat anti-rabbit IgG–Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).

    Article Snippet: After washing, the sections were incubated with goat anti-rabbit IgG - Alexa Fluor® 568 conjugate (Thermo Fisher Scientific), or goat anti-mouse IgG- Alexa Fluor® 488 conjugate, followed by incubation with DAPI to stain nuclei.

    Techniques: Expressing, Immunofluorescence, Staining, Incubation

    Validation of Annexin A1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human Annexin A1, followed by goat anti-rabbit IgG - Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).

    Journal: Scientific Reports

    Article Title: Identification of specific biomarkers for gastric adenocarcinoma by ITRAQ proteomic approach

    doi: 10.1038/srep38871

    Figure Lengend Snippet: Validation of Annexin A1 expression by immunofluorescence staining. Frozen sections from stage I and III GC ( A ) and their adjacent normal gastric tissues ( B ) were incubated with antibody against human Annexin A1, followed by goat anti-rabbit IgG - Alexa Fluor® 568 conjugate. Nuclei are stained with DAPI (blue).

    Article Snippet: After washing, the sections were incubated with goat anti-rabbit IgG - Alexa Fluor® 568 conjugate (Thermo Fisher Scientific), or goat anti-mouse IgG- Alexa Fluor® 488 conjugate, followed by incubation with DAPI to stain nuclei.

    Techniques: Expressing, Immunofluorescence, Staining, Incubation

    MAG-C42R has an altered subcellular localization. ( A ) The localization of EGFP-tagged MAG clones in HEK293T cells. EGFP was directly visualized. The presence of MAG in the endoplasmic reticulum and Golgi complex was analyzed by probing cells with anti-calnexin and anti-GM130 antibodies, respectively, both detected with Alexa Fluor 568-conjugated secondary antibodies (red). DNA was stained with Hoechst 33342 (blue). Photographs were acquired using a Zeiss Axio Imager Z1 microscope. Bars: 10 μm. ( B ) The co-immunoprecipitation of EGFP-tagged MAG clones with endogenous calnexin and GM130 in HEK293T cells. Cell lysates were immunoprecipitated with anti-GFP antibody. The immunoprecipitates were subjected to immunoblotting with anti-GM130 antibody, then stripped and reprobed with anti-calnexin antibody, followed by stripping and reprobing with anti-EGFP antibody. The same methodology was applied to whole cell lysates. Calnexin and GM130 immunoblotting figures have different exposure times (original blots are presented in  Figure S2 ).

    Journal: Journal of Clinical Medicine

    Article Title: Novel MAG Variant Causes Cerebellar Ataxia with Oculomotor Apraxia: Molecular Basis and Expanded Clinical Phenotype

    doi: 10.3390/jcm9041212

    Figure Lengend Snippet: MAG-C42R has an altered subcellular localization. ( A ) The localization of EGFP-tagged MAG clones in HEK293T cells. EGFP was directly visualized. The presence of MAG in the endoplasmic reticulum and Golgi complex was analyzed by probing cells with anti-calnexin and anti-GM130 antibodies, respectively, both detected with Alexa Fluor 568-conjugated secondary antibodies (red). DNA was stained with Hoechst 33342 (blue). Photographs were acquired using a Zeiss Axio Imager Z1 microscope. Bars: 10 μm. ( B ) The co-immunoprecipitation of EGFP-tagged MAG clones with endogenous calnexin and GM130 in HEK293T cells. Cell lysates were immunoprecipitated with anti-GFP antibody. The immunoprecipitates were subjected to immunoblotting with anti-GM130 antibody, then stripped and reprobed with anti-calnexin antibody, followed by stripping and reprobing with anti-EGFP antibody. The same methodology was applied to whole cell lysates. Calnexin and GM130 immunoblotting figures have different exposure times (original blots are presented in Figure S2 ).

    Article Snippet: Secondary antibodies: horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (sc-2005, Santa Cruz Biotechnology, Heidelberg, Germany), HRP-conjugated goat anti-rabbit IgG (401393, Calbiochem, EDM Millipore, Darmstadt, Germany,), goat anti-mouse IgG Alexa Fluor® 568 conjugate (A-11004, ThermoFisher Scientific, Waltham, MA, USA), and goat anti-rabbit IgG Alexa Fluor® 568 conjugate (A-11011, ThermoFisher Scientific, Waltham, MA, USA).

    Techniques: Clone Assay, Staining, Microscopy, Immunoprecipitation, Stripping Membranes

    Twinfilin regulates capping protein localization and dynamics. (A) Localization of EGFP-CP in wild-type and twf1/twf2-KO B16-F1 cells, where F-actin was visualized with AlexaFluor-568 phalloidin. Panels in the middle and right are magnifications of lamellipodial regions highlighted in the whole cell images in left. Scale bars = 10 μ m. (B) Examples of line profiles generated across the center of lamellipodia as indicated with dotted lines. Data represent mean of 5 measurements of individual lamellipodia, with standard deviations shown. The ‘0 μ m’ value in x-axis is set to correspond the peak intensity of phalloidin. (C) The ratio of CP and F-actin co-localization widths were detected by measuring the width of localization at 50% of maximum intensity. Data points represent measurements from individual lamellipodia with mean values and standard deviations shown. Statistical significance was calculated with Student’s unpaired, two-tailed t-test. ****, p

    Journal: bioRxiv

    Article Title: Twinfilin uncaps filament barbed ends to promote turnover of lamellipodial actin networks

    doi: 10.1101/864769

    Figure Lengend Snippet: Twinfilin regulates capping protein localization and dynamics. (A) Localization of EGFP-CP in wild-type and twf1/twf2-KO B16-F1 cells, where F-actin was visualized with AlexaFluor-568 phalloidin. Panels in the middle and right are magnifications of lamellipodial regions highlighted in the whole cell images in left. Scale bars = 10 μ m. (B) Examples of line profiles generated across the center of lamellipodia as indicated with dotted lines. Data represent mean of 5 measurements of individual lamellipodia, with standard deviations shown. The ‘0 μ m’ value in x-axis is set to correspond the peak intensity of phalloidin. (C) The ratio of CP and F-actin co-localization widths were detected by measuring the width of localization at 50% of maximum intensity. Data points represent measurements from individual lamellipodia with mean values and standard deviations shown. Statistical significance was calculated with Student’s unpaired, two-tailed t-test. ****, p

    Article Snippet: Other antibodies used in the study were: Rabbit anti-twinfilin-2 antibody (Sigma-Aldrich #HPA053874, WB, 1:100), rabbit anti-CAPZß antibody (Sigma-Aldrich, #HPA031531, WB, 1:100), mouse anti-α-tubulin antibody (Sigma-Aldrich, #T5168, WB 1:10,000), mouse anti-ß-actin antibody (Sigma-Aldrich, #A5441, WB, 1:10,000), Rabbit anti-p34-Arc/ARPC2 (Merck Millipore, #07-227, dilution in immunofluorescence (IF), 1:200), goat anti-Rabbit IgG AlexaFluor-488 conjugated secondary antibody (Thermo Fisher, #A-11034, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-568 conjugated secondary antibody (Thermo Fisher, #A-11011, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-647 conjugated secondary antibody (Thermo Fisher, #A-32733, IF, 1:400), goat anti-Mouse IgG HRP conjugated secondary antibody (Thermo Fisher, #31430, WB, 1:10,000), goat anti-Rabbit IgG HRP conjugated secondary antibody (Thermo Fisher, #32460, WB, 1:1,000).

    Techniques: Generated, Two Tailed Test

    Examples of high-content and lamellipodia protrusion analysis. (A) Representative images of segmentation procedure in high-content image analysis. Wild-type and twinfilin-deficient B16-F1 cells were stained with DAPI and CellMask Deep Red to segment nuclei (outlined with blue) and cytoplasm (outlined with yellow). The Arp2/3-complex positive structures were detected with anti-p34 antibody staining and used as a mask for the Arp2/3-complex positive F-actin structures (the most-right panel). F-actin was stained with AlexaFluor-568 phalloidin. Cells touching the border of images were excluded from analysis. (B) A representative example of twf1/twf2-KO cell migrating on laminin coated glass imaged with DIC. Direction of migration is indicated with an arrow and kymographs were generated with line drawn across the lamellipodium as indicated with dotted red line. (C) Representative examples of kymographs generated from DIC time-lapse images of wild-type, twf1/twf2 knockout, and EGFP-TWF-1 rescue cells. Protrusions velocities were measured from the overall cell front protrusion as indicated with dotted red lines.

    Journal: bioRxiv

    Article Title: Twinfilin uncaps filament barbed ends to promote turnover of lamellipodial actin networks

    doi: 10.1101/864769

    Figure Lengend Snippet: Examples of high-content and lamellipodia protrusion analysis. (A) Representative images of segmentation procedure in high-content image analysis. Wild-type and twinfilin-deficient B16-F1 cells were stained with DAPI and CellMask Deep Red to segment nuclei (outlined with blue) and cytoplasm (outlined with yellow). The Arp2/3-complex positive structures were detected with anti-p34 antibody staining and used as a mask for the Arp2/3-complex positive F-actin structures (the most-right panel). F-actin was stained with AlexaFluor-568 phalloidin. Cells touching the border of images were excluded from analysis. (B) A representative example of twf1/twf2-KO cell migrating on laminin coated glass imaged with DIC. Direction of migration is indicated with an arrow and kymographs were generated with line drawn across the lamellipodium as indicated with dotted red line. (C) Representative examples of kymographs generated from DIC time-lapse images of wild-type, twf1/twf2 knockout, and EGFP-TWF-1 rescue cells. Protrusions velocities were measured from the overall cell front protrusion as indicated with dotted red lines.

    Article Snippet: Other antibodies used in the study were: Rabbit anti-twinfilin-2 antibody (Sigma-Aldrich #HPA053874, WB, 1:100), rabbit anti-CAPZß antibody (Sigma-Aldrich, #HPA031531, WB, 1:100), mouse anti-α-tubulin antibody (Sigma-Aldrich, #T5168, WB 1:10,000), mouse anti-ß-actin antibody (Sigma-Aldrich, #A5441, WB, 1:10,000), Rabbit anti-p34-Arc/ARPC2 (Merck Millipore, #07-227, dilution in immunofluorescence (IF), 1:200), goat anti-Rabbit IgG AlexaFluor-488 conjugated secondary antibody (Thermo Fisher, #A-11034, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-568 conjugated secondary antibody (Thermo Fisher, #A-11011, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-647 conjugated secondary antibody (Thermo Fisher, #A-32733, IF, 1:400), goat anti-Mouse IgG HRP conjugated secondary antibody (Thermo Fisher, #31430, WB, 1:10,000), goat anti-Rabbit IgG HRP conjugated secondary antibody (Thermo Fisher, #32460, WB, 1:1,000).

    Techniques: Staining, Migration, Generated, Knock-Out

    Representative images of wild-type and twinfilin knockout B16-F1 cells. (A) B16-F1 cells were stained with AlexaFluor-568 phalloidin (F-actin) and anti-p34 antibody (the Arp2/3 complex). Scale bars = 10 μ M. (B) Mean F-actin intensity in B16-F1 wild-type and twinfilin knockout cells. Number of measured cells were: B16-F1 wt = 1,958, twf1-KO-g1 = 1,045, twf2-KO-g3#1 = 1,285, twf1/2-KO-g3 = 1,265, twf1/2-KO-g4 = 1,707. (C) Mean F-actin intensity in the Arp2/3 complex positive regions of B16-F1 wild-type and knockout cells. The Arp2/3 complex positive regions were identified based on p34-antibody staining. Number of measured cells were: B16-F1 wt = 1,658, twf1-KO-g1 = 884, twf2-KO-g3#1 = 1,009, twf1/2-KO-g3 = 1,100, twf1/2-KO-g4 = 1157. Statistical significances in panels B and C were calculated with Mann-Whitney two-tailed test. ****, p

    Journal: bioRxiv

    Article Title: Twinfilin uncaps filament barbed ends to promote turnover of lamellipodial actin networks

    doi: 10.1101/864769

    Figure Lengend Snippet: Representative images of wild-type and twinfilin knockout B16-F1 cells. (A) B16-F1 cells were stained with AlexaFluor-568 phalloidin (F-actin) and anti-p34 antibody (the Arp2/3 complex). Scale bars = 10 μ M. (B) Mean F-actin intensity in B16-F1 wild-type and twinfilin knockout cells. Number of measured cells were: B16-F1 wt = 1,958, twf1-KO-g1 = 1,045, twf2-KO-g3#1 = 1,285, twf1/2-KO-g3 = 1,265, twf1/2-KO-g4 = 1,707. (C) Mean F-actin intensity in the Arp2/3 complex positive regions of B16-F1 wild-type and knockout cells. The Arp2/3 complex positive regions were identified based on p34-antibody staining. Number of measured cells were: B16-F1 wt = 1,658, twf1-KO-g1 = 884, twf2-KO-g3#1 = 1,009, twf1/2-KO-g3 = 1,100, twf1/2-KO-g4 = 1157. Statistical significances in panels B and C were calculated with Mann-Whitney two-tailed test. ****, p

    Article Snippet: Other antibodies used in the study were: Rabbit anti-twinfilin-2 antibody (Sigma-Aldrich #HPA053874, WB, 1:100), rabbit anti-CAPZß antibody (Sigma-Aldrich, #HPA031531, WB, 1:100), mouse anti-α-tubulin antibody (Sigma-Aldrich, #T5168, WB 1:10,000), mouse anti-ß-actin antibody (Sigma-Aldrich, #A5441, WB, 1:10,000), Rabbit anti-p34-Arc/ARPC2 (Merck Millipore, #07-227, dilution in immunofluorescence (IF), 1:200), goat anti-Rabbit IgG AlexaFluor-488 conjugated secondary antibody (Thermo Fisher, #A-11034, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-568 conjugated secondary antibody (Thermo Fisher, #A-11011, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-647 conjugated secondary antibody (Thermo Fisher, #A-32733, IF, 1:400), goat anti-Mouse IgG HRP conjugated secondary antibody (Thermo Fisher, #31430, WB, 1:10,000), goat anti-Rabbit IgG HRP conjugated secondary antibody (Thermo Fisher, #32460, WB, 1:1,000).

    Techniques: Knock-Out, Staining, MANN-WHITNEY, Two Tailed Test

    Knockout of twinfilins leads to abnormal F-actin accumulation in lamellipodia and perinuclear region. (A) Representative images of wild-type and twf1/twf2-KO mouse B16-F1 cells stained with AlexaFluor-568 phalloidin and anti-p34 antibody to visualize F-actin and the Arp2/3 complex, respectively. Scale bar = 10 μ m. (B) F-actin intensities in the cytoplasmic regions of wild-type, twf1/twf2-KO, and knockout cells expressing EGFP-TWF-1 measured by high-content image analysis. Number of cells analyzed were: B16-F1 wt = 4,875, twf1/twf2-KO-g3 = 5,731, twf1/twf2-KO-g3 + EGFP-TWF-1 = 197. (C) Lamellipodia protrusion velocities of wild-type, twf1/twf-2 knockout, and knockout cells expressing EGFP-TWF-1. Data represent individual cells with mean and standard deviations shown. Statistical significances in panels B and D were calculated with Mann-Whitney two-tailed test. ****, p

    Journal: bioRxiv

    Article Title: Twinfilin uncaps filament barbed ends to promote turnover of lamellipodial actin networks

    doi: 10.1101/864769

    Figure Lengend Snippet: Knockout of twinfilins leads to abnormal F-actin accumulation in lamellipodia and perinuclear region. (A) Representative images of wild-type and twf1/twf2-KO mouse B16-F1 cells stained with AlexaFluor-568 phalloidin and anti-p34 antibody to visualize F-actin and the Arp2/3 complex, respectively. Scale bar = 10 μ m. (B) F-actin intensities in the cytoplasmic regions of wild-type, twf1/twf2-KO, and knockout cells expressing EGFP-TWF-1 measured by high-content image analysis. Number of cells analyzed were: B16-F1 wt = 4,875, twf1/twf2-KO-g3 = 5,731, twf1/twf2-KO-g3 + EGFP-TWF-1 = 197. (C) Lamellipodia protrusion velocities of wild-type, twf1/twf-2 knockout, and knockout cells expressing EGFP-TWF-1. Data represent individual cells with mean and standard deviations shown. Statistical significances in panels B and D were calculated with Mann-Whitney two-tailed test. ****, p

    Article Snippet: Other antibodies used in the study were: Rabbit anti-twinfilin-2 antibody (Sigma-Aldrich #HPA053874, WB, 1:100), rabbit anti-CAPZß antibody (Sigma-Aldrich, #HPA031531, WB, 1:100), mouse anti-α-tubulin antibody (Sigma-Aldrich, #T5168, WB 1:10,000), mouse anti-ß-actin antibody (Sigma-Aldrich, #A5441, WB, 1:10,000), Rabbit anti-p34-Arc/ARPC2 (Merck Millipore, #07-227, dilution in immunofluorescence (IF), 1:200), goat anti-Rabbit IgG AlexaFluor-488 conjugated secondary antibody (Thermo Fisher, #A-11034, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-568 conjugated secondary antibody (Thermo Fisher, #A-11011, IF, 1:400), goat anti-Rabbit IgG AlexaFluor-647 conjugated secondary antibody (Thermo Fisher, #A-32733, IF, 1:400), goat anti-Mouse IgG HRP conjugated secondary antibody (Thermo Fisher, #31430, WB, 1:10,000), goat anti-Rabbit IgG HRP conjugated secondary antibody (Thermo Fisher, #32460, WB, 1:1,000).

    Techniques: Knock-Out, Staining, Expressing, MANN-WHITNEY, Two Tailed Test