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P syk

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P-Syk is a phospho-specific antibody that recognizes the phosphorylated form of the Syk (spleen tyrosine kinase) protein. Syk is a key signaling molecule involved in various cellular processes, including immune response, cell adhesion, and receptor signaling. The P-Syk antibody can be used to detect and quantify the phosphorylated state of Syk in biological samples.

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Lab products found in correlation

Subcellular protein fraction kit, by Thermo Fisher Scientific (2 mentions) P stat5, by Abcam (2 mentions) P akt, by Abcam (2 mentions) P jak2, by Abcam (2 mentions) Beta2 integrin, by Abcam (2 mentions) Gm csf receptor beta common chain, by Santa Cruz Biotechnology (2 mentions) Kindlin 3, by Merck Group (2 mentions) P erk, by Abcam (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) β actin, by Abcam (2 mentions) Lsm 700 confocal microscope, by Zeiss (2 mentions) Cd3 17a2, by BioLegend (1 mentions) Axiovert camera, by Zeiss (1 mentions) Goat anti rabbit igg fitc, by Santa Cruz Biotechnology (1 mentions) Dapi counterstain, by Vector Laboratories (1 mentions) Axioscope 40 microscope camera system, by Zeiss (1 mentions) Donkey anti goat igg pe, by Santa Cruz Biotechnology (1 mentions) Cd45 microbeads, by Miltenyi Biotec (1 mentions) Protease inhibitor mix, by Merck Group (1 mentions) P akt ser473, by Cell Signaling Technology (1 mentions)

4 protocols using p syk

1

Histological and Immunohistochemical Analysis of Liver

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For histological analysis, liver specimens were fixed with 10% buffered formalin, dehydrated in ethanol, and then embedded with paraffin and stained with hematoxylin-eosin (H&E) or Masson trichrome. Percent cell death was determined on a computerized grid as we have described10 (link). For immunohistochemical analysis, slides were stained for anti-mouse CD45 (30-F11; BD Biosciences), MPO (polyclonal, Abcam), CD3 (17A2; Biolegend), CD68 (pAB; Abcam), p-Syk (polyclonal, Abcam), as well as anti-human Mincle (AT16E3, Abcam), SAP-130 (polyclonal, Abcam), and p-Syk (polyclonal, Abam). TUNEL staining was performed using a kit (EMD Millipore, Billerica, MA). For immunofluorescent imaging, murine CD45+ NPCs were isolated using CD45 MicroBeads (Miltenyi, Bergisch Gladbach, Germany) stained for Mincle (CLEC-4E polyclonal, Santa Cruz Biotechnology, Dallas, TX) or p-Syk (polyclonal, Abcam), and then detected with donkey anti-goat IgG-PE, goat anti-rabbit IgG-Fitc (both Santa Cruz Biotechnology), and DAPI counterstain (Vector Laboratories, Burlingame, CA). Light microscopic images were captured with a Zeiss Axioscope 40 microscope/camera system (Zeiss, Thornwood, NY). Immunofluorescent imaging was performed using a LSM 700 confocal microscope and an Axiovert camera (Zeiss). Data was quantified by examining 10 high powered fields (HPFs) per slide.
Greco S.H., Torres-Hernandez A., Kalabin A., Whiteman C., Rokosh R., Ravirala S., Ochi A., Gutierrez J., Salyana M.A., Mani V.R., Nagaraj S.V., Deutsch M., Seifert L., Daley D., Barilla R., Hundeyin M., Nikifrov Y., Tejada K., Gelb B.E., Katz S.C, & Miller G. (2016). Mincle Signaling Promotes Con-A Hepatitis. Journal of immunology (Baltimore, Md. : 1950), 197(7), 2816-2827.
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2

Subcellular Protein Fractionation and Immunoblotting of Dendritic Cells

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DCs were lysed in 1% Tx-100, 150 mM NaCl, 50 mM Tris pH 7.4, 10 mM ethylenediamine tetraacetic acid (EDTA), 50 mM NaF, and 1 mM sodium orthovanadate, containing protease inhibitor cocktail (Roche). Alternatively, DCs were fractionated into subcellular fractions using a subcellular protein fraction kit (Thermo Scientific, NH, USA) according to the manufacturer’s instructions. Primary antibodies against the following proteins were used (Cell Signaling, unless otherwise stated): p-JAK2, p-STAT5, p-Syk, p-p38, p-Erk, p-Akt, Akt, Src, Syk, beta-actin, beta2-integrin (Abcam, MA, USA), GM-CSF receptor beta-common chain (Santa Cruz Biotechnology, CA, USA), kindlin-3 (Sigma-Aldrich, MO, USA), talin, CD9 (BD Biosciences). After incubation in horseradish peroxidase-conjugated secondary antibodies, blots were developed by standard chemiluminescence techniques. All uncropped blots are shown in Supplementary Figure 7.
Morrison V.L., James M.J., Grzes K., Cook P., Glass D.G., Savinko T., Lek H.S., Gawden-Bone C., Watts C., Millington O.R., MacDonald A.S, & Fagerholm S.C. (2014). Loss of beta2-integrin-mediated cytoskeletal linkage reprograms dendritic cells to a mature migratory phenotype. Nature communications, 5, 5359.
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3

Subcellular Protein Fractionation and Immunoblotting of Dendritic Cells

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DCs were lysed in 1% Tx-100, 150 mM NaCl, 50 mM Tris pH 7.4, 10 mM ethylenediamine tetraacetic acid (EDTA), 50 mM NaF, and 1 mM sodium orthovanadate, containing protease inhibitor cocktail (Roche). Alternatively, DCs were fractionated into subcellular fractions using a subcellular protein fraction kit (Thermo Scientific, NH, USA) according to the manufacturer’s instructions. Primary antibodies against the following proteins were used (Cell Signaling, unless otherwise stated): p-JAK2, p-STAT5, p-Syk, p-p38, p-Erk, p-Akt, Akt, Src, Syk, beta-actin, beta2-integrin (Abcam, MA, USA), GM-CSF receptor beta-common chain (Santa Cruz Biotechnology, CA, USA), kindlin-3 (Sigma-Aldrich, MO, USA), talin, CD9 (BD Biosciences). After incubation in horseradish peroxidase-conjugated secondary antibodies, blots were developed by standard chemiluminescence techniques. All uncropped blots are shown in Supplementary Figure 7.
Morrison V.L., James M.J., Grzes K., Cook P., Glass D.G., Savinko T., Lek H.S., Gawden-Bone C., Watts C., Millington O.R., MacDonald A.S, & Fagerholm S.C. (2014). Loss of beta2-integrin-mediated cytoskeletal linkage reprograms dendritic cells to a mature migratory phenotype. Nature communications, 5, 5359.
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4

Immunoblotting of Signaling Pathways

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Cells were starved for 1 h and then stimulated for the indicated times at 37°C with 10 μg/ml anti-mouse IgM F(ab′)2 antibody (Dianova). Stimulation was stopped with cold PBS and cells were lysed for immunoblot analysis with buffer containing 20 mM Hepes (pH 7.6), 2 mM MgCl2, 150 mM NaCl, 10% glycerol, 0.1% NP40, 1 mM Na3VO4, 1 mM PMSF, and protease inhibitor mix (Sigma-Aldrich). Protein extracts corresponding to equal cell numbers were loaded onto the SDS-PAGE gel. The samples were then blotted with the following antibodies to H3 (rabbit polyclonal, Abcam); p-Syk (Tyr525/526; rabbit monoclonal; Cell Signaling Technology); p-Lyn (Tyr507; rabbit polyclonal; Cell Signaling Technology); p-Akt (Ser473; rabbit monoclonal; Cell Signaling Technology); and p-44/42 (Erk1/2; Thr202/Tyr204; rabbit monoclonal; Cell Signaling Technology). For Notch2, ILK, Grb2, and FAK immunoblots, cells were lysed as detailed above and samples were blotted with Notch2 antibody (D76A6; rabbit monoclonal; Cell Signaling Technology), ILK antibody (4G9; rabbit monoclonal; Cell Signaling Technology), and Grb2 antibody (Y237; rabbit monoclonal; Abcam), respectively, and Tubulin-HRP (Mouse IgG2b, Proteintech). Immunoblots were developed with Chemidoc Imaging System (BioRad) and analyzed with Image Lab software (BioRad).
Andreani V., Ramamoorthy S., Fässler R, & Grosschedl R. (2022). Integrin β1 regulates marginal zone B cell differentiation and PI3K signaling. The Journal of Experimental Medicine, 220(1), e20220342.
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