snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    snap surface 649 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "An examination of the divergent spatiotemporal signaling of GLP-1R versus GIPR in pancreatic beta cells"

    Article Title: An examination of the divergent spatiotemporal signaling of GLP-1R versus GIPR in pancreatic beta cells

    Journal: bioRxiv

    doi: 10.1101/2022.08.17.504231

    ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.

    Techniques Used: Western Blot

    snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface 649 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "An examination of the divergent spatiotemporal signaling of GLP-1R versus GIPR in pancreatic beta cells"

    Article Title: An examination of the divergent spatiotemporal signaling of GLP-1R versus GIPR in pancreatic beta cells

    Journal: bioRxiv

    doi: 10.1101/2022.08.17.504231

    ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.

    Techniques Used: Western Blot

    snap surface649  (New England Biolabs)


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    New England Biolabs snap surface649
    a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with <t>Surf649</t> (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.
    Snap Surface649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface649 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Multivalent interactions essential for lentiviral integrase function"

    Article Title: Multivalent interactions essential for lentiviral integrase function

    Journal: Nature Communications

    doi: 10.1038/s41467-022-29928-8

    a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with Surf649 (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with Surf649 (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.

    Techniques Used: Incubation, Microscopy, Concentration Assay, Fluorescence

    snap surface 649 fluorescent substrate  (New England Biolabs)


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    New England Biolabs snap surface 649 fluorescent substrate
    Snap Surface 649 Fluorescent Substrate, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649 fluorescent substrate/product/New England Biolabs
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    snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface 649 - by Bioz Stars, 2023-01
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    snap surface649  (New England Biolabs)


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    New England Biolabs snap surface649
    (a) Schematic of the photobleaching experiment. Intasomes containing biotinylated and Cy3-labeled vDNA were immobilized on streptavidin-coated glass cover slips. Following incubation with <t>Surf649-labeled</t> LEDGF/p75 and a wash with buffer supplemented with 0.2 - 1 M NaCl, intasomes were observed by TIRF microscopy enabling the individual steps of Surf649 photobleaching during illumination with a 640 nm laser (hν) to be counted. (b) Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. (c) Left: Examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta) and 1.0 M (dark grey). The vertical axis represents fluorescence in arbitrary units (a.u.). Right: Tukey (box-and-whiskers) plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see for further details). Each box encloses 50% of the data with the median value displayed as a horizontal line. Outliers are indicated with closed circles.
    Snap Surface649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface649 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Multivalent interactions essential for lentiviral integrase function"

    Article Title: Multivalent interactions essential for lentiviral integrase function

    Journal: bioRxiv

    doi: 10.1101/2022.01.26.477893

    (a) Schematic of the photobleaching experiment. Intasomes containing biotinylated and Cy3-labeled vDNA were immobilized on streptavidin-coated glass cover slips. Following incubation with Surf649-labeled LEDGF/p75 and a wash with buffer supplemented with 0.2 - 1 M NaCl, intasomes were observed by TIRF microscopy enabling the individual steps of Surf649 photobleaching during illumination with a 640 nm laser (hν) to be counted. (b) Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. (c) Left: Examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta) and 1.0 M (dark grey). The vertical axis represents fluorescence in arbitrary units (a.u.). Right: Tukey (box-and-whiskers) plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see for further details). Each box encloses 50% of the data with the median value displayed as a horizontal line. Outliers are indicated with closed circles.
    Figure Legend Snippet: (a) Schematic of the photobleaching experiment. Intasomes containing biotinylated and Cy3-labeled vDNA were immobilized on streptavidin-coated glass cover slips. Following incubation with Surf649-labeled LEDGF/p75 and a wash with buffer supplemented with 0.2 - 1 M NaCl, intasomes were observed by TIRF microscopy enabling the individual steps of Surf649 photobleaching during illumination with a 640 nm laser (hν) to be counted. (b) Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. (c) Left: Examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta) and 1.0 M (dark grey). The vertical axis represents fluorescence in arbitrary units (a.u.). Right: Tukey (box-and-whiskers) plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see for further details). Each box encloses 50% of the data with the median value displayed as a horizontal line. Outliers are indicated with closed circles.

    Techniques Used: Labeling, Incubation, Microscopy, Concentration Assay, Fluorescence


    Figure Legend Snippet:

    Techniques Used:

    snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    photostable fluorescent substrates snap surface 649  (New England Biolabs)


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    New England Biolabs photostable fluorescent substrates snap surface 649
    Photostable Fluorescent Substrates Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
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    snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
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    snap surface 649  (New England Biolabs)


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    New England Biolabs snap surface 649
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649/product/New England Biolabs
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    New England Biolabs snap surface 649
    ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.
    Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94
    New England Biolabs snap surface649
    a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with <t>Surf649</t> (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.
    Snap Surface649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface649 - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    New England Biolabs snap surface 649 fluorescent substrate
    a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with <t>Surf649</t> (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.
    Snap Surface 649 Fluorescent Substrate, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 649 fluorescent substrate/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface 649 fluorescent substrate - by Bioz Stars, 2023-01
    94/100 stars
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    94
    New England Biolabs photostable fluorescent substrates snap surface 649
    a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with <t>Surf649</t> (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.
    Photostable Fluorescent Substrates Snap Surface 649, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/photostable fluorescent substrates snap surface 649/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    photostable fluorescent substrates snap surface 649 - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.

    Journal: bioRxiv

    Article Title: An examination of the divergent spatiotemporal signaling of GLP-1R versus GIPR in pancreatic beta cells

    doi: 10.1101/2022.08.17.504231

    Figure Lengend Snippet: ( A ) Western blot assessment of SNAP-GLP-1R or GIPR over tubulin levels in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for 6 hrs in the presence of the protein synthesis inhibitor cycloheximide; n =3. ( B ) Representative Western blot results from (A). Note that the top bands were used to quantify the SNAP-receptor levels, as they correspond to the glycosylated forms of the receptors, known to be biologically active and correctly inserted at the plasma membrane . ( C ) Percentage of GLP-1R vs GIPR, labelled with the cell-permeable SNAP-tag probe BG-OG, and corresponding representative images from INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells with or without stimulation with 100 nM GLP-1 or GIP, respectively, for the indicated times in the presence of cycloheximide; n =4. ( D ) Percentage of co-localization (Mander’s coefficient) and representative images of SNAP-GLP-1R vs -GIPR (labelled with SNAP-Surface 649) with Lysotracker Green in INS-1 832/3 SNAP-GLP-1R or SNAP-GIPR cells stimulated with 100 nM GLP-1 or GIP for 1 hr; n =5. Data are mean ± SEM, compared by ratio-paired or unpaired t-test or two-way ANOVA with Sidak’s post-hoc test; ***p<0.001, ****p<0.0001.

    Article Snippet: INS-1 832/3 SNAP- GLP- 1R or SNAP-GIPR cells were labelled in suspension with 40 nM SNAP- Lumi4- Tb and 1 mM SNAP- Surface 649 (New England Biolabs, Hitchin, UK) for 1 hr at room temperature in complete medium.

    Techniques: Western Blot

    a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with Surf649 (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Multivalent interactions essential for lentiviral integrase function

    doi: 10.1038/s41467-022-29928-8

    Figure Lengend Snippet: a Schematic of the photobleaching experiment. Intasomes containing biotin- and Cy3-conjugated vDNA were immobilized on a streptavidin-coated cover slip (light blue rectangle). IN and vDNA are shown as gray and black cartoons, respectively; biotin and Cy3 are depicted as a purple circle and yellow solar symbol, respectively. Following incubation with LEDGF/p75 (red cartoons) conjugated with Surf649 (red solar symbol) and a wash with buffer containing 0.2, 0.5, or 1 M NaCl, immobilized intasomes were observed by TIRF microscopy. The individual steps of Surf649 photobleaching during illumination with a 640-nm laser (hν) were counted. b Representative images of surface attached intasome (vDNA-Cy3, yellow; LEDGF/p75-Surf649, red) molecules. The field of view of 14.4 by 14.4 μm shown here is representative of the dataset, which included three areas of 81.9 by 81.9 μm (see Methods section for details). Dotted, white line squares indicate individual intasome-LEDGF/p75 complexes. c Left: examples of stepwise photobleaching traces of LEDGF/p75-Surf649 at increasing NaCl concentration: 0.2 M (blue), 0.5 M (magenta), and 1.0 M (dark gray). The vertical axis represents fluorescence in arbitrary units (arb. units). Right: Box-and-whiskers plots summarizing statistical analysis of the number of LEDGF/p75-Surf649 photobleaching steps per intasome, for various NaCl concentrations (see Supplementary Table for further details). Each box encloses data between 25th and 75th percentiles, with the median value displayed as a horizontal line. Vertical lines (whiskers) indicate 10th and 90th percentiles; outliers are indicated with closed circles. Source data are provided as a Source Data file.

    Article Snippet: LEGDF-SNAPf protein was labeled with SNAP-Surface649 (Surf649, New England Biolabs).

    Techniques: Incubation, Microscopy, Concentration Assay, Fluorescence