For immunofluorescence of endogenous adhesion proteins, cells were incubated with opsonized particles, fixed for 10 minutes at 37˚C, then rinsed twice and fixed with 4% paraformaldehyde (Electron microscopy science) in cytoskeleton buffer (10 mM MES pH 6.1, 138 mM KCl, 3 mM MgCl2, 2 mM EGTA) for 20 minutes. Free aldehydes were quenched 100 mM glycine for 10 minutes, and cells were permeabilized with 0.5% Triton X-100 for 5 minutes. Endogenous proteins were immuno-labelled using the following antibodies: anti α-Actinin (Sigma-Aldrich A5044, 1:500 dilution), anti Vinculin (Sigma-Aldrich V9131, 1:500 dilution), anti Zyxin (B71, 1:600 dilution), anti phosphotyrosine (EMD Millipore 05-321, 1:300 dilution), anti phospho-FAK Y397 (Invitrogen 44722G, 1:100 dilution), anti phospho-Paxillin Y31 (Invitrogen 44720G, 1:100 dilution), anti phospho-Paxillin Y118 (Invitrogen 44722G, 1:100 dilution), anti Syk Y348 (BD Pharmingen 558167, 1:200 dilution). Cy3-conjugated secondary antibodies (Jackson) and Alexa Fluor 488 phalloidin (Fischer scientific) were used at 1:400 dilution. Samples were mounted on glass slides with Dako (Agilent).
Anti syk y348
Manufactured by BD
Anti Syk Y348 is a laboratory reagent that targets the phosphorylation of the tyrosine 348 residue on the Syk protein. It can be used in various in vitro research applications to study the role of Syk signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (2 mentions)
Cy3 conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions)
Anti vinculin, by Merck Group (2 mentions)
Anti phosphotyrosine, by Merck Group (2 mentions)
Anti α actinin, by Merck Group (2 mentions)
Anti zyxin, by Merck Group (2 mentions)
Anti phospho fak y397, by Thermo Fisher Scientific (2 mentions)
Anti phospho paxillin y31, by Thermo Fisher Scientific (2 mentions)
Anti phospho paxillin y118, by Thermo Fisher Scientific (2 mentions)
2 protocols using anti syk y348
1
Immunofluorescence of Adhesion Proteins
2
Immunofluorescence of Adhesion Proteins
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For immunofluorescence of endogenous adhesion proteins, cells were incubated with opsonized particles, fixed for 10 minutes at 37˚C, then rinsed twice and fixed with 4% paraformaldehyde (Electron microscopy science) in cytoskeleton buffer (10 mM MES pH 6.1, 138 mM KCl, 3 mM MgCl2, 2 mM EGTA) for 20 minutes. Free aldehydes were quenched 100 mM glycine for 10 minutes, and cells were permeabilized with 0.5% Triton X-100 for 5 minutes. Endogenous proteins were immuno-labelled using the following antibodies: anti α-Actinin (Sigma-Aldrich A5044, 1:500 dilution), anti Vinculin (Sigma-Aldrich V9131, 1:500 dilution), anti Zyxin (B71, 1:600 dilution), anti phosphotyrosine (EMD Millipore 05-321, 1:300 dilution), anti phospho-FAK Y397 (Invitrogen 44722G, 1:100 dilution), anti phospho-Paxillin Y31 (Invitrogen 44720G, 1:100 dilution), anti phospho-Paxillin Y118 (Invitrogen 44722G, 1:100 dilution), anti Syk Y348 (BD Pharmingen 558167, 1:200 dilution). Cy3-conjugated secondary antibodies (Jackson) and Alexa Fluor 488 phalloidin (Fischer scientific) were used at 1:400 dilution. Samples were mounted on glass slides with Dako (Agilent).
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