active src (py418) Search Results


c src py 418 antibodies  (New England Biolabs)
96
New England Biolabs c src py 418 antibodies
C Src Py 418 Antibodies, 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
https://www.bioz.com/product/active+src+%28py418%29/pmc03314471-546-3-8?v=New+England+Biolabs
Average 96 stars, based on 1 article reviews
c src py 418 antibodies - by Bioz Stars, 2026-08
96/100 stars
  Buy from Supplier
py418 sfk  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc py418 sfk
Thy-1 modulates force-dependent <t>SFK</t> and RhoA adhesion signaling. (a) Cartoon schematic of the tensile force application assay. Magnetic beads coated with FN are allowed to bind cell surface receptors, after which a permanent magnet is used to apply tensile force for varying amounts of time. Cells are then lysed for biochemical analysis. (b) RhoA activity was measured after tensile magnetic force stimulation of FN- or anti-transferrin receptor beads bound to empty vector control (cont. vector)– and Thy-1 WT –expressing RFL-6 cells. Apparent rate constants ( K ) for a single-phase association model are indicated (mean ± SEM); plateau phase of control vector was statistically different from Thy-1 WT ( n = 6 biological replicates from two independent experiments; unpaired t test, P < 0.05). Normalized baseline level for zero force is indicated in gray. TfR indicates transferrin receptor. (c) Immunoblots of FA proteins bound to FN beads (adhesion complex) before (− force) or after (+ force) 3 min of constant force application or total cell lysates in control vector and Thy-1 WT RFL-6 cells are shown. (d) Immunoblots were quantified for n = 3–4 biological replicates from two independent experiments for association to FN-bead adhesion complexes before (− force) or after (+ force). Significance was calculated between all groups for individual adhesion components using one-way analysis of variance and Tukey’s post test. *, P < 0.05; **, P < 0.01 between indicated groups.
Py418 Sfk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/active+src+%28py418%29/pmc04602038-261-87-95?v=Cell+Signaling+Technology+Inc
Average 96 stars, based on 1 article reviews
py418 sfk - by Bioz Stars, 2026-08
96/100 stars
  Buy from Supplier

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Thy-1 modulates force-dependent SFK and RhoA adhesion signaling. (a) Cartoon schematic of the tensile force application assay. Magnetic beads coated with FN are allowed to bind cell surface receptors, after which a permanent magnet is used to apply tensile force for varying amounts of time. Cells are then lysed for biochemical analysis. (b) RhoA activity was measured after tensile magnetic force stimulation of FN- or anti-transferrin receptor beads bound to empty vector control (cont. vector)– and Thy-1 WT –expressing RFL-6 cells. Apparent rate constants ( K ) for a single-phase association model are indicated (mean ± SEM); plateau phase of control vector was statistically different from Thy-1 WT ( n = 6 biological replicates from two independent experiments; unpaired t test, P < 0.05). Normalized baseline level for zero force is indicated in gray. TfR indicates transferrin receptor. (c) Immunoblots of FA proteins bound to FN beads (adhesion complex) before (− force) or after (+ force) 3 min of constant force application or total cell lysates in control vector and Thy-1 WT RFL-6 cells are shown. (d) Immunoblots were quantified for n = 3–4 biological replicates from two independent experiments for association to FN-bead adhesion complexes before (− force) or after (+ force). Significance was calculated between all groups for individual adhesion components using one-way analysis of variance and Tukey’s post test. *, P < 0.05; **, P < 0.01 between indicated groups.

Journal: The Journal of Cell Biology

Article Title: Conformational coupling of integrin and Thy-1 regulates Fyn priming and fibroblast mechanotransduction

doi: 10.1083/jcb.201505007

Figure Lengend Snippet: Thy-1 modulates force-dependent SFK and RhoA adhesion signaling. (a) Cartoon schematic of the tensile force application assay. Magnetic beads coated with FN are allowed to bind cell surface receptors, after which a permanent magnet is used to apply tensile force for varying amounts of time. Cells are then lysed for biochemical analysis. (b) RhoA activity was measured after tensile magnetic force stimulation of FN- or anti-transferrin receptor beads bound to empty vector control (cont. vector)– and Thy-1 WT –expressing RFL-6 cells. Apparent rate constants ( K ) for a single-phase association model are indicated (mean ± SEM); plateau phase of control vector was statistically different from Thy-1 WT ( n = 6 biological replicates from two independent experiments; unpaired t test, P < 0.05). Normalized baseline level for zero force is indicated in gray. TfR indicates transferrin receptor. (c) Immunoblots of FA proteins bound to FN beads (adhesion complex) before (− force) or after (+ force) 3 min of constant force application or total cell lysates in control vector and Thy-1 WT RFL-6 cells are shown. (d) Immunoblots were quantified for n = 3–4 biological replicates from two independent experiments for association to FN-bead adhesion complexes before (− force) or after (+ force). Significance was calculated between all groups for individual adhesion components using one-way analysis of variance and Tukey’s post test. *, P < 0.05; **, P < 0.01 between indicated groups.

Article Snippet: Cultured cells directly lysed in Laemmli buffer or bead-associated adhesion complexes were heat-denatured at 95°C for 5 min. Proteins were separated by SDS-PAGE, transferred to nitrocellulose membrane, and blotted with primary antibodies for Thy-1 (1:1,000, rabbit, 9798; Cell Signaling Technology), mouse Thy-1.2 (1:2,500, rat, 53-2.1; BD), α v integrin (1:1,000, rabbit, AB1930; Millipore), α 5 integrin (1:1,000, rabbit, AB1928; Millipore), talin (1:800, mouse, 8d4; Sigma-Aldrich), paxillin (1:1,000, mouse, 5H11; Millipore), pY397-FAK (1:1,000, rabbit, 44-624G; Invitrogen), Fyn (1:1,000, mouse, 1S; Millipore), c-Src (1:1,000, rabbit, 32G6; Cell Signaling Technology), pY418-SFK (1:1,000, rabbit, 2101; or 1:1,000, rabbit, D49G4; Cell Signaling Technology), flottilin-2 (1:1,000, rabbit, L294; Cell Signaling Technology), and Cbp (1:800, mouse, PAG-C1; Pierce Chemical).

Techniques: Magnetic Beads, Activity Assay, Plasmid Preparation, Control, Expressing, Western Blot

Integrin avidity to FN is decreased by Thy-1 cis binding and modulates SFK-mediated soft ECM sensing. (a) Schematic of the single-cell force spectroscopy assay. Cells are attached to an AFM cantilever via physioadsorption of concanavalin A and then touched to a ligand-coated substrate (FN-gl) with controlled force and contact duration. The probe is then extended from the substrate, and the resultant detachment forces are measured. (b) An example force versus extension curve is shown. Individual unbinding events are observed as steps in the force trace, and the maximum detachment force, F D , is indicated. Smoothed data using the Savitsky-Golay method are shown in red. (c) F D was measured for varying contact times in RFL-6 cells expressing Thy-1 WT , Thy-1 D37E , or control vector constructs. F D versus contact time curves for ∼50 contacts in three to five individual cells were fit with a single-phase association model. Data shown are mean ± SD pooled from two independent experiments. (d) Adhesion rate (apparent rate constant, K ) decreases because of Thy-1 WT , but not in the RLD mutant. (e) RFL-6s were plated on 10 µg/ml FN-gl, fixed after 10 or 30 min, and stained for active α v β 3 (WOW-1) and active SFK (pY418-SFK) to measure integrin clustering and activity during initial cell spreading. Bar, 20 µm. (f) Integrin cluster size (black, left) and pY418-SFK (red, right) were quantified (see Materials and methods). Statistical marks (percentage for α v β 3 cluster size; P < 0.001) indicate significance with respect to Thy-1 WT at 10 min except where indicated. (g) Blocking integrin binding or SFK signaling normalizes aberrant mechanotransduction effects of Thy-1 loss on soft ECMs. Cell spreading of Thy-1 pos (red) or Thy-1 neg MLFs on soft (1.8 kPa) FN-PA treated with 10 µg/ml IgG control or function-blocking anti-α v or anti-β 3 antibodies, RGD or negative control RGE (0.5 mM), and SFK inhibitor PP2 or its negative control PP3 (10 µM) is shown. Bar, 50 µm. (h) Cell area was quantified and represented as box-and-whisker plots (10th–90th percentiles with outlier points shown). Significance with respect to IgG-treated Thy-1 neg MLFs is shown. All statistical significance was calculated using one-way analysis of variance and Tukey’s post test. *, P < 0.05; **, P < 0.01; ***, P < 0.001 between indicated groups.

Journal: The Journal of Cell Biology

Article Title: Conformational coupling of integrin and Thy-1 regulates Fyn priming and fibroblast mechanotransduction

doi: 10.1083/jcb.201505007

Figure Lengend Snippet: Integrin avidity to FN is decreased by Thy-1 cis binding and modulates SFK-mediated soft ECM sensing. (a) Schematic of the single-cell force spectroscopy assay. Cells are attached to an AFM cantilever via physioadsorption of concanavalin A and then touched to a ligand-coated substrate (FN-gl) with controlled force and contact duration. The probe is then extended from the substrate, and the resultant detachment forces are measured. (b) An example force versus extension curve is shown. Individual unbinding events are observed as steps in the force trace, and the maximum detachment force, F D , is indicated. Smoothed data using the Savitsky-Golay method are shown in red. (c) F D was measured for varying contact times in RFL-6 cells expressing Thy-1 WT , Thy-1 D37E , or control vector constructs. F D versus contact time curves for ∼50 contacts in three to five individual cells were fit with a single-phase association model. Data shown are mean ± SD pooled from two independent experiments. (d) Adhesion rate (apparent rate constant, K ) decreases because of Thy-1 WT , but not in the RLD mutant. (e) RFL-6s were plated on 10 µg/ml FN-gl, fixed after 10 or 30 min, and stained for active α v β 3 (WOW-1) and active SFK (pY418-SFK) to measure integrin clustering and activity during initial cell spreading. Bar, 20 µm. (f) Integrin cluster size (black, left) and pY418-SFK (red, right) were quantified (see Materials and methods). Statistical marks (percentage for α v β 3 cluster size; P < 0.001) indicate significance with respect to Thy-1 WT at 10 min except where indicated. (g) Blocking integrin binding or SFK signaling normalizes aberrant mechanotransduction effects of Thy-1 loss on soft ECMs. Cell spreading of Thy-1 pos (red) or Thy-1 neg MLFs on soft (1.8 kPa) FN-PA treated with 10 µg/ml IgG control or function-blocking anti-α v or anti-β 3 antibodies, RGD or negative control RGE (0.5 mM), and SFK inhibitor PP2 or its negative control PP3 (10 µM) is shown. Bar, 50 µm. (h) Cell area was quantified and represented as box-and-whisker plots (10th–90th percentiles with outlier points shown). Significance with respect to IgG-treated Thy-1 neg MLFs is shown. All statistical significance was calculated using one-way analysis of variance and Tukey’s post test. *, P < 0.05; **, P < 0.01; ***, P < 0.001 between indicated groups.

Article Snippet: Cultured cells directly lysed in Laemmli buffer or bead-associated adhesion complexes were heat-denatured at 95°C for 5 min. Proteins were separated by SDS-PAGE, transferred to nitrocellulose membrane, and blotted with primary antibodies for Thy-1 (1:1,000, rabbit, 9798; Cell Signaling Technology), mouse Thy-1.2 (1:2,500, rat, 53-2.1; BD), α v integrin (1:1,000, rabbit, AB1930; Millipore), α 5 integrin (1:1,000, rabbit, AB1928; Millipore), talin (1:800, mouse, 8d4; Sigma-Aldrich), paxillin (1:1,000, mouse, 5H11; Millipore), pY397-FAK (1:1,000, rabbit, 44-624G; Invitrogen), Fyn (1:1,000, mouse, 1S; Millipore), c-Src (1:1,000, rabbit, 32G6; Cell Signaling Technology), pY418-SFK (1:1,000, rabbit, 2101; or 1:1,000, rabbit, D49G4; Cell Signaling Technology), flottilin-2 (1:1,000, rabbit, L294; Cell Signaling Technology), and Cbp (1:800, mouse, PAG-C1; Pierce Chemical).

Techniques: Binding Assay, Force Spectroscopy, Expressing, Control, Plasmid Preparation, Construct, Mutagenesis, Staining, Activity Assay, Blocking Assay, Negative Control, Whisker Assay

A mechanistic model of Thy-1’s effects on integrin avidity and signaling. Thy-1 binds α v integrins preferentially in their inactive, “off” state, which couples integrins with membrane raft domains and their localized signaling molecules, such as Fyn and Cbp, before FN recognition (top, left). FN binding and receptor clustering enriches for these domains within growing FAs while promoting conformation-dependent integrin/SFK accessibility of binding motifs and sensitive responses to stimuli (e.g., force; top, right). In the absence of Thy-1, a higher probability of α v integrin binding to FN exists, thus eliciting elevated levels of baseline integrin signaling, potentially through c-Src recruitment and activation (bottom, left). As a result of decoupling of integrin and membrane rafts, raft-associated molecules (Fyn, Cbp) are no longer recruited to growing FAs and sensitive signaling (e.g., Cbp-dependent negative feedback on c-Src regulation and force-dependent Fyn activation) is not as prominent (bottom, right).

Journal: The Journal of Cell Biology

Article Title: Conformational coupling of integrin and Thy-1 regulates Fyn priming and fibroblast mechanotransduction

doi: 10.1083/jcb.201505007

Figure Lengend Snippet: A mechanistic model of Thy-1’s effects on integrin avidity and signaling. Thy-1 binds α v integrins preferentially in their inactive, “off” state, which couples integrins with membrane raft domains and their localized signaling molecules, such as Fyn and Cbp, before FN recognition (top, left). FN binding and receptor clustering enriches for these domains within growing FAs while promoting conformation-dependent integrin/SFK accessibility of binding motifs and sensitive responses to stimuli (e.g., force; top, right). In the absence of Thy-1, a higher probability of α v integrin binding to FN exists, thus eliciting elevated levels of baseline integrin signaling, potentially through c-Src recruitment and activation (bottom, left). As a result of decoupling of integrin and membrane rafts, raft-associated molecules (Fyn, Cbp) are no longer recruited to growing FAs and sensitive signaling (e.g., Cbp-dependent negative feedback on c-Src regulation and force-dependent Fyn activation) is not as prominent (bottom, right).

Article Snippet: Cultured cells directly lysed in Laemmli buffer or bead-associated adhesion complexes were heat-denatured at 95°C for 5 min. Proteins were separated by SDS-PAGE, transferred to nitrocellulose membrane, and blotted with primary antibodies for Thy-1 (1:1,000, rabbit, 9798; Cell Signaling Technology), mouse Thy-1.2 (1:2,500, rat, 53-2.1; BD), α v integrin (1:1,000, rabbit, AB1930; Millipore), α 5 integrin (1:1,000, rabbit, AB1928; Millipore), talin (1:800, mouse, 8d4; Sigma-Aldrich), paxillin (1:1,000, mouse, 5H11; Millipore), pY397-FAK (1:1,000, rabbit, 44-624G; Invitrogen), Fyn (1:1,000, mouse, 1S; Millipore), c-Src (1:1,000, rabbit, 32G6; Cell Signaling Technology), pY418-SFK (1:1,000, rabbit, 2101; or 1:1,000, rabbit, D49G4; Cell Signaling Technology), flottilin-2 (1:1,000, rabbit, L294; Cell Signaling Technology), and Cbp (1:800, mouse, PAG-C1; Pierce Chemical).

Techniques: Membrane, Binding Assay, Activation Assay