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  • 99
    Thermo Fisher albumin from bovine serum
    Albumin From Bovine Serum, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore dnp bsa albumin from bovine serum bsa 2 4 dinitrophenylated
    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml <t>DNP-BSA.</t> Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).
    Dnp Bsa Albumin From Bovine Serum Bsa 2 4 Dinitrophenylated, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnp bsa albumin from bovine serum bsa 2 4 dinitrophenylated/product/Millipore
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    99
    Thermo Fisher antibiotic antimycotic 100x
    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml <t>DNP-BSA.</t> Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).
    Antibiotic Antimycotic 100x, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 207 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibiotic antimycotic 100x/product/Thermo Fisher
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    97
    Thermo Fisher albumax i lipid rich bovine serum albumin thermo fisher scientific
    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml <t>DNP-BSA.</t> Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).
    Albumax I Lipid Rich Bovine Serum Albumin Thermo Fisher Scientific, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher b 27 supplement 50x minus vitamin a
    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml <t>DNP-BSA.</t> Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).
    B 27 Supplement 50x Minus Vitamin A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher goat anti rabbit igg h l secondary antibody
    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml <t>DNP-BSA.</t> Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).
    Goat Anti Rabbit Igg H L Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1787 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 1787 article reviews
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    97
    Thermo Fisher bovine serum albumin thermo fisher scientific
    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml <t>DNP-BSA.</t> Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).
    Bovine Serum Albumin Thermo Fisher Scientific, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml DNP-BSA. Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).

    Journal: Molecular Biology of the Cell

    Article Title: Differential mast cell outcomes are sensitive to FcεRI-Syk binding kinetics

    doi: 10.1091/mbc.E17-06-0350

    Figure Lengend Snippet: Syk mNG -Y130E exhibits a faster FcεRI off-rate. (A) TIRF images of AF647-IgE (magenta) and Syk mNG (green) membrane localization in Syk-KO cells reconstituted with Syk mNG -WT (left), Syk mNG -Y130E (middle), or Syk mNG -Y130F (right) after 4–5 min stimulation with 0.1 µg/ml DNP-BSA. Scale bar: 5 µm. (B) Quantification of FcεRI recruitment capacity for Syk. Individual aggregates of AF647-IgE were masked, and the total intensity within the mask for both the AF647-IgE and Syk mNG channels is plotted per aggregate. Recruitment is similar for Syk mNG -WT and each mutant. (C) Cumulative probability distributions of trajectory lengths for Syk mNG -WT, Syk mNG -Y130E, and Syk mNG -Y130F both before (dashed lines) and after (solid lines) addition of 1 µg/ml DNP-BSA. (D) Bar graph depicts the slow off-rate ( k s ) found when fitting the distributions in C. Fraction of slow off-rate component (α s ) increases with DNP-BSA dose for Syk mNG -WT and each mutant (right). Error bars are a 68% credible interval (see Materials and Methods ).

    Article Snippet: DNP-BSA containing ∼15–25 DNP per BSA was purchased from Thermo Fisher Scientific (A23018), and DNP-lysine was purchased from Sigma (Nε-DNP-l -lysine hydrochloride, D0380).

    Techniques: Mutagenesis

    Syk Recruitment to FcεRI. Syk-KO RBL cells expressing Syk mNG (green) were primed with AF647-IgE (magenta) and imaged after cross-linking with 0.1 µg/ml DNP-BSA. (A) Sample images from a confocal time series showing the redistribution of AF647-IgE-FcεRI and Syk mNG upon FcεRI stimulation (see also Supplemental Video 1). Resting cross-section shows homogeneous distribution of AF647-IgE-FcεRI at the plasma membrane and Syk mNG in the cytosol. Upon cross-linking (5 min), FcεRI aggregation and Syk mNG colocalization is readily seen at the adherent cell surface. Scale bar: 10.3 µm. White boxes in the “Overlay” panels are enlarged in the “Zoom” panels. Scale bar: 2 µm. (B) Treatment with 1 µM dasatinib results in a loss of Syk mNG recruitment (bottom) to FcεRI aggregates (top). Images of the adherent cell membrane acquired in TIRF. Scale bar: 5 µm. (C) Both FcεRI and Syk mNG aggregates (top) are disrupted upon addition of 100 mM monovalent DNP-lysine (middle). Scale bar: 5 µm. Selected images from a time series (bottom) show the dispersion of an individual Syk aggregate within seconds of DNP-lysine addition. Individual Syk mNG molecules can be seen diffusing away from the original diffraction-limited aggregate (see also Supplemental Video 2). Images acquired in TIRF. Scale bar: 1 µm. (D) Plot of positive correlation between Syk mNG and AF647-IgE intensity within each AF647-IgE aggregate. (E) Selected images from a confocal time series before and after photobleaching (at t = 0 s) of an individual Syk mNG aggregate. Scale bar: 1 µm. Bottom curve quantifies the rapid recovery of mNG fluorescence intensity within the bleached region (white circles).

    Journal: Molecular Biology of the Cell

    Article Title: Differential mast cell outcomes are sensitive to FcεRI-Syk binding kinetics

    doi: 10.1091/mbc.E17-06-0350

    Figure Lengend Snippet: Syk Recruitment to FcεRI. Syk-KO RBL cells expressing Syk mNG (green) were primed with AF647-IgE (magenta) and imaged after cross-linking with 0.1 µg/ml DNP-BSA. (A) Sample images from a confocal time series showing the redistribution of AF647-IgE-FcεRI and Syk mNG upon FcεRI stimulation (see also Supplemental Video 1). Resting cross-section shows homogeneous distribution of AF647-IgE-FcεRI at the plasma membrane and Syk mNG in the cytosol. Upon cross-linking (5 min), FcεRI aggregation and Syk mNG colocalization is readily seen at the adherent cell surface. Scale bar: 10.3 µm. White boxes in the “Overlay” panels are enlarged in the “Zoom” panels. Scale bar: 2 µm. (B) Treatment with 1 µM dasatinib results in a loss of Syk mNG recruitment (bottom) to FcεRI aggregates (top). Images of the adherent cell membrane acquired in TIRF. Scale bar: 5 µm. (C) Both FcεRI and Syk mNG aggregates (top) are disrupted upon addition of 100 mM monovalent DNP-lysine (middle). Scale bar: 5 µm. Selected images from a time series (bottom) show the dispersion of an individual Syk aggregate within seconds of DNP-lysine addition. Individual Syk mNG molecules can be seen diffusing away from the original diffraction-limited aggregate (see also Supplemental Video 2). Images acquired in TIRF. Scale bar: 1 µm. (D) Plot of positive correlation between Syk mNG and AF647-IgE intensity within each AF647-IgE aggregate. (E) Selected images from a confocal time series before and after photobleaching (at t = 0 s) of an individual Syk mNG aggregate. Scale bar: 1 µm. Bottom curve quantifies the rapid recovery of mNG fluorescence intensity within the bleached region (white circles).

    Article Snippet: DNP-BSA containing ∼15–25 DNP per BSA was purchased from Thermo Fisher Scientific (A23018), and DNP-lysine was purchased from Sigma (Nε-DNP-l -lysine hydrochloride, D0380).

    Techniques: Expressing, Fluorescence

    FcεRI-Syk off-rate is independent of antigen dose or aggregate size. (A) AF647-IgE–labeled RBL cells cross-linked for 5 min with indicated DNP-BSA concentration and imaged (left) at the adherent surface in TIRF (scale bar: 5 µm) and using (right) dSTORM (scale bar: 2 µm). (B) Clustering of localizations in dSTORM images from A using a hierarchical clustering algorithm (Matlab, MathWorks). Cluster sizes are shown as gray dots, and the distribution is summarized by the mean (red cross), the median (red line), and the 25th and 75th percentiles (blue box). Kolmogorov-Smirnov tests show significant ( p

    Journal: Molecular Biology of the Cell

    Article Title: Differential mast cell outcomes are sensitive to FcεRI-Syk binding kinetics

    doi: 10.1091/mbc.E17-06-0350

    Figure Lengend Snippet: FcεRI-Syk off-rate is independent of antigen dose or aggregate size. (A) AF647-IgE–labeled RBL cells cross-linked for 5 min with indicated DNP-BSA concentration and imaged (left) at the adherent surface in TIRF (scale bar: 5 µm) and using (right) dSTORM (scale bar: 2 µm). (B) Clustering of localizations in dSTORM images from A using a hierarchical clustering algorithm (Matlab, MathWorks). Cluster sizes are shown as gray dots, and the distribution is summarized by the mean (red cross), the median (red line), and the 25th and 75th percentiles (blue box). Kolmogorov-Smirnov tests show significant ( p

    Article Snippet: DNP-BSA containing ∼15–25 DNP per BSA was purchased from Thermo Fisher Scientific (A23018), and DNP-lysine was purchased from Sigma (Nε-DNP-l -lysine hydrochloride, D0380).

    Techniques: Labeling, Concentration Assay

    Key mast cell outcomes are impaired in Syk mNG -Y130E cells. (A) Degranulation measured by relative β-hexosaminidase released after 30 min of incubation with 0.1 µg/ml DNP-BSA in Syk-KO cells reconstituted with Syk mNG -WT, Syk mNG -Y130E, or Syk mNG -Y130F. (B) Comparison of cytokine concentration in cell media of Syk-KO cells reconstituted with Syk mNG -WT, Syk mNG -Y130E, or Syk mNG -Y130F before and after 3-h stimulation with 0.1 µg/ml DNP-BSA. Results were repeated at least three times for all four cytokines. Bar plots represent mean and SD of technical replicates in one representative sample preparation. (C) The formation of actin plaques at the basolateral surface (left) and ruffling on the apical surface (right) in response to 0.1 µg/ml DNP-BSA treatment in both Syk mNG -WT and Syk mNG -Y130E reconstituted Syk-KO cells. Filamentous actin labeled with phalloidin-AF647 (red). Scale bar: 2 µm. (D, E) Heat maps of relative changes in intracellular calcium concentration upon either addition of a (D) low dose (0.001 µg/ml) or (E) high dose (1 µg/ml) of DNP-BSA. Each row represents the ratio of Fura-2 emission using 350-nm/380-nm excitation for a single cell over time. Ratio color bar scale range, one- to twofold increase: blue–red. (F) Relative increase in Fura-2 ratio per cell after addition. (G) Time between antigen addition and response for each cell. (F,G) Stimulated with 1 µg/ml DNP-BSA and calculated as described in Materials and Methods .

    Journal: Molecular Biology of the Cell

    Article Title: Differential mast cell outcomes are sensitive to FcεRI-Syk binding kinetics

    doi: 10.1091/mbc.E17-06-0350

    Figure Lengend Snippet: Key mast cell outcomes are impaired in Syk mNG -Y130E cells. (A) Degranulation measured by relative β-hexosaminidase released after 30 min of incubation with 0.1 µg/ml DNP-BSA in Syk-KO cells reconstituted with Syk mNG -WT, Syk mNG -Y130E, or Syk mNG -Y130F. (B) Comparison of cytokine concentration in cell media of Syk-KO cells reconstituted with Syk mNG -WT, Syk mNG -Y130E, or Syk mNG -Y130F before and after 3-h stimulation with 0.1 µg/ml DNP-BSA. Results were repeated at least three times for all four cytokines. Bar plots represent mean and SD of technical replicates in one representative sample preparation. (C) The formation of actin plaques at the basolateral surface (left) and ruffling on the apical surface (right) in response to 0.1 µg/ml DNP-BSA treatment in both Syk mNG -WT and Syk mNG -Y130E reconstituted Syk-KO cells. Filamentous actin labeled with phalloidin-AF647 (red). Scale bar: 2 µm. (D, E) Heat maps of relative changes in intracellular calcium concentration upon either addition of a (D) low dose (0.001 µg/ml) or (E) high dose (1 µg/ml) of DNP-BSA. Each row represents the ratio of Fura-2 emission using 350-nm/380-nm excitation for a single cell over time. Ratio color bar scale range, one- to twofold increase: blue–red. (F) Relative increase in Fura-2 ratio per cell after addition. (G) Time between antigen addition and response for each cell. (F,G) Stimulated with 1 µg/ml DNP-BSA and calculated as described in Materials and Methods .

    Article Snippet: DNP-BSA containing ∼15–25 DNP per BSA was purchased from Thermo Fisher Scientific (A23018), and DNP-lysine was purchased from Sigma (Nε-DNP-l -lysine hydrochloride, D0380).

    Techniques: Incubation, Concentration Assay, Sample Prep, Labeling

    Quantifying FcεRI-Syk interactions using single-particle tracking of Syk mNG . Single Syk mNG molecules were tracked using TIRF microscopy as they bound and dissociated from the membrane. (A) Example of Syk mNG trajectories detected at the plasma membrane (left). Projection of trajectories in the time dimension (middle) shows the trajectory lengths. Enlargement of a single trajectory that lasts ∼10 s (right). (B) Cumulative probability distribution of Syk mNG trajectory lengths before (black) and 4–5 min after addition of 1 µg/ml DNP-BSA (green). Solid gray lines represent the fit to the data. (C) Bar graph depicts fast and slow off-rates found when fitting the distributions in B. See Materials and Methods for details. (D) Fraction of the slow off-rate component (α s ) increases from ∼5% to 23% after addition of 1 µg/ml DNP-BSA. Treatment with 1 µM dasatinib (Das) reduces α s in both resting and activated cells. Error bars in C and D are a 68% credible interval as described in Materials and Methods . See also Supplemental Table S1.

    Journal: Molecular Biology of the Cell

    Article Title: Differential mast cell outcomes are sensitive to FcεRI-Syk binding kinetics

    doi: 10.1091/mbc.E17-06-0350

    Figure Lengend Snippet: Quantifying FcεRI-Syk interactions using single-particle tracking of Syk mNG . Single Syk mNG molecules were tracked using TIRF microscopy as they bound and dissociated from the membrane. (A) Example of Syk mNG trajectories detected at the plasma membrane (left). Projection of trajectories in the time dimension (middle) shows the trajectory lengths. Enlargement of a single trajectory that lasts ∼10 s (right). (B) Cumulative probability distribution of Syk mNG trajectory lengths before (black) and 4–5 min after addition of 1 µg/ml DNP-BSA (green). Solid gray lines represent the fit to the data. (C) Bar graph depicts fast and slow off-rates found when fitting the distributions in B. See Materials and Methods for details. (D) Fraction of the slow off-rate component (α s ) increases from ∼5% to 23% after addition of 1 µg/ml DNP-BSA. Treatment with 1 µM dasatinib (Das) reduces α s in both resting and activated cells. Error bars in C and D are a 68% credible interval as described in Materials and Methods . See also Supplemental Table S1.

    Article Snippet: DNP-BSA containing ∼15–25 DNP per BSA was purchased from Thermo Fisher Scientific (A23018), and DNP-lysine was purchased from Sigma (Nε-DNP-l -lysine hydrochloride, D0380).

    Techniques: Single-particle Tracking, Microscopy

    Phosphorylation kinetics of Syk and downstream signaling partners. (A) Western blot detection of the Syk phosphorylation profile in Syk-KO cells reconstituted with Syk mNG -WT, Syk mNG -Y130E, or Syk mNG -Y130F in response to stimulation with 0.1 µg/ml DNP-BSA for 5 min. Bar plots (right) represent mean and SD of relative Syk phosphorylation level from at least three experiments. Phosphorylation of tyrosines associated with Syk autocatalytic activity (pY519/520 and pY342) is significantly reduced ( t test: *, p

    Journal: Molecular Biology of the Cell

    Article Title: Differential mast cell outcomes are sensitive to FcεRI-Syk binding kinetics

    doi: 10.1091/mbc.E17-06-0350

    Figure Lengend Snippet: Phosphorylation kinetics of Syk and downstream signaling partners. (A) Western blot detection of the Syk phosphorylation profile in Syk-KO cells reconstituted with Syk mNG -WT, Syk mNG -Y130E, or Syk mNG -Y130F in response to stimulation with 0.1 µg/ml DNP-BSA for 5 min. Bar plots (right) represent mean and SD of relative Syk phosphorylation level from at least three experiments. Phosphorylation of tyrosines associated with Syk autocatalytic activity (pY519/520 and pY342) is significantly reduced ( t test: *, p

    Article Snippet: DNP-BSA containing ∼15–25 DNP per BSA was purchased from Thermo Fisher Scientific (A23018), and DNP-lysine was purchased from Sigma (Nε-DNP-l -lysine hydrochloride, D0380).

    Techniques: Western Blot, Activity Assay