snap surface alexa fluor 647  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647
    Snap Surface Alexa Fluor 647, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    s9136s  (New England Biolabs)


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    New England Biolabs s9136s

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    1) Product Images from "Molecular dissection of PI3Kβ synergistic activation by receptor tyrosine kinases, GβGγ, and Rho-family GTPases"

    Article Title: Molecular dissection of PI3Kβ synergistic activation by receptor tyrosine kinases, GβGγ, and Rho-family GTPases

    Journal: eLife

    doi: 10.7554/eLife.88991


    Figure Legend Snippet:

    Techniques Used: Transfection, Protease Inhibitor, Recombinant, Sequencing, Expressing, Plasmid Preparation, Software, Cell Culture

    snap surface alexa fluor 647  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647
    Snap Surface Alexa Fluor 647, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    snap surface alexa fluor 647 dye  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647 dye
    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa <t>Fluor</t> <t>647</t> (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
    Snap Surface Alexa Fluor 647 Dye, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "MRAP2 modifies the signaling and oligomerization state of the melanocortin-4 receptor"

    Article Title: MRAP2 modifies the signaling and oligomerization state of the melanocortin-4 receptor

    Journal: bioRxiv

    doi: 10.1101/2024.04.09.588099

    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa Fluor 647 (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
    Figure Legend Snippet: (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa Fluor 647 (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).

    Techniques Used: Microscopy, Membrane, Expressing, Labeling, Transfection, Fluorescence, Concentration Assay

    snap surface alexa fluor 647 neb  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647 neb
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    snap surface alexa fluor 647  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647
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    snap surface af647  (New England Biolabs)


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    snap surface alexa fluor 647  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647
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    snap surface alexa flour 647  (New England Biolabs)


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    New England Biolabs snap surface alexa flour 647
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    snap surface alexa 647  (New England Biolabs)


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    New England Biolabs snap surface alexa 647
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    snap surface alexa fluor 647  (New England Biolabs)


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    New England Biolabs snap surface alexa fluor 647
    (A) Purified CENP-T and Ndc80 constructs used in this study were analyzed by SDS-PAGE. (B) Example photobleaching curves for GFP-tagged CENP-T 6D (C, D) Histograms of integral intensities collected from photobleaching curves. Binned data are represented with mean ± SEM, red lines are fittings of the main distributions with Gaussian functions. Peaks of intensities close to zero represent the background values. Panel (C) shows integral intensities of GFP fluorophore, number of independent experiments (N) = 3, total number of analyzed dots (n) = 47. Panel (D) shows integral intensities of Alexa <t>Fluor</t> <t>647</t> dye, N = 4, n = 112. (E-I) Histograms of the number of GFP molecules per fluorescent dot in the chamber with indicated GFP-tagged CENP-T construct, N = 3, n > 100 per protein. Values indicated on the graphs correspond to the peak value determined with Gaussian fitting and represent the number of CENP-T molecules per dot. Dots with several GFP molecules are rare and likely represent two or more molecules localizing close together.
    Snap Surface Alexa Fluor 647, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Molecular density-accelerated binding-site maturation underlies CENP-T-dependent kinetochore assembly"

    Article Title: Molecular density-accelerated binding-site maturation underlies CENP-T-dependent kinetochore assembly

    Journal: bioRxiv

    doi: 10.1101/2024.02.25.581584

    (A) Purified CENP-T and Ndc80 constructs used in this study were analyzed by SDS-PAGE. (B) Example photobleaching curves for GFP-tagged CENP-T 6D (C, D) Histograms of integral intensities collected from photobleaching curves. Binned data are represented with mean ± SEM, red lines are fittings of the main distributions with Gaussian functions. Peaks of intensities close to zero represent the background values. Panel (C) shows integral intensities of GFP fluorophore, number of independent experiments (N) = 3, total number of analyzed dots (n) = 47. Panel (D) shows integral intensities of Alexa Fluor 647 dye, N = 4, n = 112. (E-I) Histograms of the number of GFP molecules per fluorescent dot in the chamber with indicated GFP-tagged CENP-T construct, N = 3, n > 100 per protein. Values indicated on the graphs correspond to the peak value determined with Gaussian fitting and represent the number of CENP-T molecules per dot. Dots with several GFP molecules are rare and likely represent two or more molecules localizing close together.
    Figure Legend Snippet: (A) Purified CENP-T and Ndc80 constructs used in this study were analyzed by SDS-PAGE. (B) Example photobleaching curves for GFP-tagged CENP-T 6D (C, D) Histograms of integral intensities collected from photobleaching curves. Binned data are represented with mean ± SEM, red lines are fittings of the main distributions with Gaussian functions. Peaks of intensities close to zero represent the background values. Panel (C) shows integral intensities of GFP fluorophore, number of independent experiments (N) = 3, total number of analyzed dots (n) = 47. Panel (D) shows integral intensities of Alexa Fluor 647 dye, N = 4, n = 112. (E-I) Histograms of the number of GFP molecules per fluorescent dot in the chamber with indicated GFP-tagged CENP-T construct, N = 3, n > 100 per protein. Values indicated on the graphs correspond to the peak value determined with Gaussian fitting and represent the number of CENP-T molecules per dot. Dots with several GFP molecules are rare and likely represent two or more molecules localizing close together.

    Techniques Used: Purification, Construct, SDS Page

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    New England Biolabs snap surface alexa fluor 647
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    New England Biolabs snap surface alexa fluor 647 dye
    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa <t>Fluor</t> <t>647</t> (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
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    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa <t>Fluor</t> <t>647</t> (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
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    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa <t>Fluor</t> <t>647</t> (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
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    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa <t>Fluor</t> <t>647</t> (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
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    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa <t>Fluor</t> <t>647</t> (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).
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    Image Search Results


    Journal: eLife

    Article Title: Molecular dissection of PI3Kβ synergistic activation by receptor tyrosine kinases, GβGγ, and Rho-family GTPases

    doi: 10.7554/eLife.88991

    Figure Lengend Snippet:

    Article Snippet: Chemical compound, drug , SNAP-Surface Alexa Fluor 647 , NEB , S9136S , .

    Techniques: Transfection, Protease Inhibitor, Recombinant, Sequencing, Expressing, Plasmid Preparation, Software, Cell Culture

    (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa Fluor 647 (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).

    Journal: bioRxiv

    Article Title: MRAP2 modifies the signaling and oligomerization state of the melanocortin-4 receptor

    doi: 10.1101/2024.04.09.588099

    Figure Lengend Snippet: (A) Representative images from TIRF microscopy of the basolateral membrane of HEK293AD cells expressing SNAP-tagged MC4R labeled using Alexa Fluor 647 (gray) and Nb35-eYFP (green). Images were taken pre-(top) and 4 min post-(bottom) stimulation with 1 µM NDP-α-MSH for cells expressing SNAP-MC4R alone or with MRAP2 (at a ratio of 1:4) and indicate recruitment of Nb35 (green intensity). (B) Kinetics of recruitment of Nb35 to the SNAP-MC4R–Gs complex, without (black; 5 transfections, 51 single cells) and with MRAP2 (1+4) (turquoise; 5 transfections, 55 cells). Shown are the normalized fluorescence intensities recorded at the membrane in the eYFP channel; the shaded area represents SEM. (C) Mean concentration-response curves of Rluc8-miniGs recruitment to the plasma membrane localization sensor rGFP-CAAX upon MC4R stimulation with α-MSH for 45 minutes, in the presence or absence of MRAP2 or RAMP3, at the indicated ratio with respect to MC4R, in transiently transfected HEK293-SL cells. Normalized data are expressed as mean ± SEM of three independent experiments. (D) EC50 values from the miniGs recruitment BRET experiments. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with Dunnett’s multiple comparisons post-hoc test (*** = p < 0.001).

    Article Snippet: Next day, transfected cells were labeled with SNAP-Surface Alexa Fluor 647 dye (NEB) as described earlier.

    Techniques: Microscopy, Membrane, Expressing, Labeling, Transfection, Fluorescence, Concentration Assay