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sarafluor650b  (GORYO Chemical)


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    GORYO Chemical sarafluor650b
    sEVs bind to laminin on the plasma membranes of living MRC-5 cells, as revealed by pseudo-real-time super-resolution movie observation. (A) Schematic diagram for generating merged movies of dSTORM images of the ECM structures and single-particle images of sEVs. Data acquisition was performed by observing single fluorescent molecules of <t>SF650B</t> at 200 frames/s (3,504 frames), and dSTORM images were reconstructed using the data acquired every 5.0 s (=1,002 frames). The first dSTORM image was reconstructed using frames 1-1,002 and the process was then repeated by shifting the initial frames backward by 6 frames; thus, 417 dSTORM images were obtained. These dSTORM still images were connected to construct the “dSTORM movie”. The single-particle movies of sEVs were subjected to a rolling-average for 6 frames and synchronously merged with the dSTORM movie. (B-D) Conventional immunofluorescence images (top-left) and dSTORM images (bottom-left) of the ECM structures (B: fibronectin, C: collagen type I, D: laminin) on the cells. dSTORM images of the ECM structures (magenta) and single-particle images of sEV-CD63Halo7-TMR particles (green) were merged (right). sEVs localized near (<100 nm) the boundary of ECM structures and sEVs localized alone are indicated by yellow and white arrowheads, respectively. (E) The image sequence (every 0.3 sec) of laminin obtained by dSTORM (magenta) and a single sEV-CD63Halo7-TMR particle (green) on the living MRC-5 cell membrane. sEVs colocalized with laminin, as indicated by the yellow arrowhead. (F) Colocalization analysis method. The normalized relative frequency was defined as the ratio of the average value of the spatial pair correlation function of the actual image to that of randomly distributed spots generated by a computer. Zero on the x-axis indicates the contour of the ECM structures in the dSTORM images determined by the kernel density estimation (KDE) method. When sEVs are enriched near the ECM structures, the normalized relative frequency is greater than 1. (G) Probability density analysis of the sEVs and ECM structures. The colored areas indicate regions within the ECM structures. The sEV-CD63Halo7-TMR particles localized near the contour of laminin ( n = 20 cells).
    Sarafluor650b, supplied by GORYO Chemical, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sarafluor+650b-nhs/SaraFluor+650B-NHS/bio_rxiv__2024__04__11__589011-365-15-17
    Average 94 stars, based on 4 article reviews
    sarafluor650b - by Bioz Stars, 2026-10
    94/100 stars

    Images

    1) Product Images from "Extracellular vesicles adhere to cells predominantly through the interaction of CD151-associated integrin heterodimers and GM1 with laminin"

    Article Title: Extracellular vesicles adhere to cells predominantly through the interaction of CD151-associated integrin heterodimers and GM1 with laminin

    Journal: bioRxiv

    doi: 10.1101/2024.04.11.589011

    sEVs bind to laminin on the plasma membranes of living MRC-5 cells, as revealed by pseudo-real-time super-resolution movie observation. (A) Schematic diagram for generating merged movies of dSTORM images of the ECM structures and single-particle images of sEVs. Data acquisition was performed by observing single fluorescent molecules of SF650B at 200 frames/s (3,504 frames), and dSTORM images were reconstructed using the data acquired every 5.0 s (=1,002 frames). The first dSTORM image was reconstructed using frames 1-1,002 and the process was then repeated by shifting the initial frames backward by 6 frames; thus, 417 dSTORM images were obtained. These dSTORM still images were connected to construct the “dSTORM movie”. The single-particle movies of sEVs were subjected to a rolling-average for 6 frames and synchronously merged with the dSTORM movie. (B-D) Conventional immunofluorescence images (top-left) and dSTORM images (bottom-left) of the ECM structures (B: fibronectin, C: collagen type I, D: laminin) on the cells. dSTORM images of the ECM structures (magenta) and single-particle images of sEV-CD63Halo7-TMR particles (green) were merged (right). sEVs localized near (<100 nm) the boundary of ECM structures and sEVs localized alone are indicated by yellow and white arrowheads, respectively. (E) The image sequence (every 0.3 sec) of laminin obtained by dSTORM (magenta) and a single sEV-CD63Halo7-TMR particle (green) on the living MRC-5 cell membrane. sEVs colocalized with laminin, as indicated by the yellow arrowhead. (F) Colocalization analysis method. The normalized relative frequency was defined as the ratio of the average value of the spatial pair correlation function of the actual image to that of randomly distributed spots generated by a computer. Zero on the x-axis indicates the contour of the ECM structures in the dSTORM images determined by the kernel density estimation (KDE) method. When sEVs are enriched near the ECM structures, the normalized relative frequency is greater than 1. (G) Probability density analysis of the sEVs and ECM structures. The colored areas indicate regions within the ECM structures. The sEV-CD63Halo7-TMR particles localized near the contour of laminin ( n = 20 cells).
    Figure Legend Snippet: sEVs bind to laminin on the plasma membranes of living MRC-5 cells, as revealed by pseudo-real-time super-resolution movie observation. (A) Schematic diagram for generating merged movies of dSTORM images of the ECM structures and single-particle images of sEVs. Data acquisition was performed by observing single fluorescent molecules of SF650B at 200 frames/s (3,504 frames), and dSTORM images were reconstructed using the data acquired every 5.0 s (=1,002 frames). The first dSTORM image was reconstructed using frames 1-1,002 and the process was then repeated by shifting the initial frames backward by 6 frames; thus, 417 dSTORM images were obtained. These dSTORM still images were connected to construct the “dSTORM movie”. The single-particle movies of sEVs were subjected to a rolling-average for 6 frames and synchronously merged with the dSTORM movie. (B-D) Conventional immunofluorescence images (top-left) and dSTORM images (bottom-left) of the ECM structures (B: fibronectin, C: collagen type I, D: laminin) on the cells. dSTORM images of the ECM structures (magenta) and single-particle images of sEV-CD63Halo7-TMR particles (green) were merged (right). sEVs localized near (<100 nm) the boundary of ECM structures and sEVs localized alone are indicated by yellow and white arrowheads, respectively. (E) The image sequence (every 0.3 sec) of laminin obtained by dSTORM (magenta) and a single sEV-CD63Halo7-TMR particle (green) on the living MRC-5 cell membrane. sEVs colocalized with laminin, as indicated by the yellow arrowhead. (F) Colocalization analysis method. The normalized relative frequency was defined as the ratio of the average value of the spatial pair correlation function of the actual image to that of randomly distributed spots generated by a computer. Zero on the x-axis indicates the contour of the ECM structures in the dSTORM images determined by the kernel density estimation (KDE) method. When sEVs are enriched near the ECM structures, the normalized relative frequency is greater than 1. (G) Probability density analysis of the sEVs and ECM structures. The colored areas indicate regions within the ECM structures. The sEV-CD63Halo7-TMR particles localized near the contour of laminin ( n = 20 cells).

    Techniques Used: Single Particle, Construct, Immunofluorescence, Sequencing, Membrane, Generated

    Related Articles

    Purification:

    Article Title: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX.
    Article Snippet: ubated for 14 h at 25 °C. The remainder of the protein purification protocol was identical to that for Imp β1–mEosEM. The spontaneously blinking dye HMSiR maleimide (SaraFluor 650B-maleimide; A209-01, Goryo Chemical) was attached to the C-terminal cysteine on the anti-GFP nanobody LaG-9(S151C) by incubating with a 15-fold molar excess at room temperature for 15 min to yield NbGFP–HMSiR. Imp α was u

    Protein Purification:

    Article Title: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX.
    Article Snippet: ubated for 14 h at 25 °C. The remainder of the protein purification protocol was identical to that for Imp β1–mEosEM. The spontaneously blinking dye HMSiR maleimide (SaraFluor 650B-maleimide; A209-01, Goryo Chemical) was attached to the C-terminal cysteine on the anti-GFP nanobody LaG-9(S151C) by incubating with a 15-fold molar excess at room temperature for 15 min to yield NbGFP–HMSiR. Imp α was u

    Incubation:

    Article Title: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX.
    Article Snippet: ubated for 14 h at 25 °C. The remainder of the protein purification protocol was identical to that for Imp β1–mEosEM. The spontaneously blinking dye HMSiR maleimide (SaraFluor 650B-maleimide; A209-01, Goryo Chemical) was attached to the C-terminal cysteine on the anti-GFP nanobody LaG-9(S151C) by incubating with a 15-fold molar excess at room temperature for 15 min to yield NbGFP–HMSiR. Imp α was u

    Imaging:

    Article Title: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX.
    Article Snippet: ubated for 14 h at 25 °C. The remainder of the protein purification protocol was identical to that for Imp β1–mEosEM. The spontaneously blinking dye HMSiR maleimide (SaraFluor 650B-maleimide; A209-01, Goryo Chemical) was attached to the C-terminal cysteine on the anti-GFP nanobody LaG-9(S151C) by incubating with a 15-fold molar excess at room temperature for 15 min to yield NbGFP–HMSiR. Imp α was u

    Fluorescence:

    Article Title: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX.
    Article Snippet: ubated for 14 h at 25 °C. The remainder of the protein purification protocol was identical to that for Imp β1–mEosEM. The spontaneously blinking dye HMSiR maleimide (SaraFluor 650B-maleimide; A209-01, Goryo Chemical) was attached to the C-terminal cysteine on the anti-GFP nanobody LaG-9(S151C) by incubating with a 15-fold molar excess at room temperature for 15 min to yield NbGFP–HMSiR. Imp α was u

    Chromatography:

    Article Title: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX.
    Article Snippet: ubated for 14 h at 25 °C. The remainder of the protein purification protocol was identical to that for Imp β1–mEosEM. The spontaneously blinking dye HMSiR maleimide (SaraFluor 650B-maleimide; A209-01, Goryo Chemical) was attached to the C-terminal cysteine on the anti-GFP nanobody LaG-9(S151C) by incubating with a 15-fold molar excess at room temperature for 15 min to yield NbGFP–HMSiR. Imp α was u



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    sEVs bind to laminin on the plasma membranes of living MRC-5 cells, as revealed by pseudo-real-time super-resolution movie observation. (A) Schematic diagram for generating merged movies of dSTORM images of the ECM structures and single-particle images of sEVs. Data acquisition was performed by observing single fluorescent molecules of <t>SF650B</t> at 200 frames/s (3,504 frames), and dSTORM images were reconstructed using the data acquired every 5.0 s (=1,002 frames). The first dSTORM image was reconstructed using frames 1-1,002 and the process was then repeated by shifting the initial frames backward by 6 frames; thus, 417 dSTORM images were obtained. These dSTORM still images were connected to construct the “dSTORM movie”. The single-particle movies of sEVs were subjected to a rolling-average for 6 frames and synchronously merged with the dSTORM movie. (B-D) Conventional immunofluorescence images (top-left) and dSTORM images (bottom-left) of the ECM structures (B: fibronectin, C: collagen type I, D: laminin) on the cells. dSTORM images of the ECM structures (magenta) and single-particle images of sEV-CD63Halo7-TMR particles (green) were merged (right). sEVs localized near (<100 nm) the boundary of ECM structures and sEVs localized alone are indicated by yellow and white arrowheads, respectively. (E) The image sequence (every 0.3 sec) of laminin obtained by dSTORM (magenta) and a single sEV-CD63Halo7-TMR particle (green) on the living MRC-5 cell membrane. sEVs colocalized with laminin, as indicated by the yellow arrowhead. (F) Colocalization analysis method. The normalized relative frequency was defined as the ratio of the average value of the spatial pair correlation function of the actual image to that of randomly distributed spots generated by a computer. Zero on the x-axis indicates the contour of the ECM structures in the dSTORM images determined by the kernel density estimation (KDE) method. When sEVs are enriched near the ECM structures, the normalized relative frequency is greater than 1. (G) Probability density analysis of the sEVs and ECM structures. The colored areas indicate regions within the ECM structures. The sEV-CD63Halo7-TMR particles localized near the contour of laminin ( n = 20 cells).
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    Image Search Results


    sEVs bind to laminin on the plasma membranes of living MRC-5 cells, as revealed by pseudo-real-time super-resolution movie observation. (A) Schematic diagram for generating merged movies of dSTORM images of the ECM structures and single-particle images of sEVs. Data acquisition was performed by observing single fluorescent molecules of SF650B at 200 frames/s (3,504 frames), and dSTORM images were reconstructed using the data acquired every 5.0 s (=1,002 frames). The first dSTORM image was reconstructed using frames 1-1,002 and the process was then repeated by shifting the initial frames backward by 6 frames; thus, 417 dSTORM images were obtained. These dSTORM still images were connected to construct the “dSTORM movie”. The single-particle movies of sEVs were subjected to a rolling-average for 6 frames and synchronously merged with the dSTORM movie. (B-D) Conventional immunofluorescence images (top-left) and dSTORM images (bottom-left) of the ECM structures (B: fibronectin, C: collagen type I, D: laminin) on the cells. dSTORM images of the ECM structures (magenta) and single-particle images of sEV-CD63Halo7-TMR particles (green) were merged (right). sEVs localized near (<100 nm) the boundary of ECM structures and sEVs localized alone are indicated by yellow and white arrowheads, respectively. (E) The image sequence (every 0.3 sec) of laminin obtained by dSTORM (magenta) and a single sEV-CD63Halo7-TMR particle (green) on the living MRC-5 cell membrane. sEVs colocalized with laminin, as indicated by the yellow arrowhead. (F) Colocalization analysis method. The normalized relative frequency was defined as the ratio of the average value of the spatial pair correlation function of the actual image to that of randomly distributed spots generated by a computer. Zero on the x-axis indicates the contour of the ECM structures in the dSTORM images determined by the kernel density estimation (KDE) method. When sEVs are enriched near the ECM structures, the normalized relative frequency is greater than 1. (G) Probability density analysis of the sEVs and ECM structures. The colored areas indicate regions within the ECM structures. The sEV-CD63Halo7-TMR particles localized near the contour of laminin ( n = 20 cells).

    Journal: bioRxiv

    Article Title: Extracellular vesicles adhere to cells predominantly through the interaction of CD151-associated integrin heterodimers and GM1 with laminin

    doi: 10.1101/2024.04.11.589011

    Figure Lengend Snippet: sEVs bind to laminin on the plasma membranes of living MRC-5 cells, as revealed by pseudo-real-time super-resolution movie observation. (A) Schematic diagram for generating merged movies of dSTORM images of the ECM structures and single-particle images of sEVs. Data acquisition was performed by observing single fluorescent molecules of SF650B at 200 frames/s (3,504 frames), and dSTORM images were reconstructed using the data acquired every 5.0 s (=1,002 frames). The first dSTORM image was reconstructed using frames 1-1,002 and the process was then repeated by shifting the initial frames backward by 6 frames; thus, 417 dSTORM images were obtained. These dSTORM still images were connected to construct the “dSTORM movie”. The single-particle movies of sEVs were subjected to a rolling-average for 6 frames and synchronously merged with the dSTORM movie. (B-D) Conventional immunofluorescence images (top-left) and dSTORM images (bottom-left) of the ECM structures (B: fibronectin, C: collagen type I, D: laminin) on the cells. dSTORM images of the ECM structures (magenta) and single-particle images of sEV-CD63Halo7-TMR particles (green) were merged (right). sEVs localized near (<100 nm) the boundary of ECM structures and sEVs localized alone are indicated by yellow and white arrowheads, respectively. (E) The image sequence (every 0.3 sec) of laminin obtained by dSTORM (magenta) and a single sEV-CD63Halo7-TMR particle (green) on the living MRC-5 cell membrane. sEVs colocalized with laminin, as indicated by the yellow arrowhead. (F) Colocalization analysis method. The normalized relative frequency was defined as the ratio of the average value of the spatial pair correlation function of the actual image to that of randomly distributed spots generated by a computer. Zero on the x-axis indicates the contour of the ECM structures in the dSTORM images determined by the kernel density estimation (KDE) method. When sEVs are enriched near the ECM structures, the normalized relative frequency is greater than 1. (G) Probability density analysis of the sEVs and ECM structures. The colored areas indicate regions within the ECM structures. The sEV-CD63Halo7-TMR particles localized near the contour of laminin ( n = 20 cells).

    Article Snippet: For immunostaining of the ECM components for dSTORM movie observation, 0.67 μl of 5.6 μg/ml SaraFluor650B (SF650B, Goryo Chemical) NHS ester was incubated with 100 μl of 1 mg/ml anti-rabbit IgG antibody (0212-0081, Cappel) in 0.1 M NaHCO 3 for 60 min at room temperature.

    Techniques: Single Particle, Construct, Immunofluorescence, Sequencing, Membrane, Generated