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Mouse Monoclonal to Syk

SYK-01 (IgG1)

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Background

Syk is a cytoplasmic protein tyrosine kinase that translocates to the plasma membrane upon B cell antigen receptor (BCR) or the high-affinity IgE receptor (FcepsilonRI) triggering, and phosphorylates downstream adaptor proteins, thereby providing docking sites for initiation of subsequent signaling pathways, such as calcium mobilization, cytoskeleton remodeling, or transcription of specific genes. Syk binds to the receptor assemblies through interactions of its pair of SH2 domains with ITAM motives of the receptor, which have been phosphorylated by Src-family kinases. These kinases also help to activate Syk by phosphorylation of its activation loop.

Specificity

The antibody SYK-01 reacts with protein tyrosine kinase p72Syk (Syk; Syk family tyrosine-specific phospho-transferase), which is required for the transduction of signals through the B cell antigen receptor (BCR) and the high affinity IgE receptor (FcepsilonRI).

Regulatory Status

Immunogen

Recombinant fragment (aa 5-360) of human Syk.

Species Reactivity:

  • Human
  • Mouse
  • Rat
  • Other not tested

Negative Species:

Applications:

  • Immunoprecipitation
  • Western Blotting
    Recommended dilution:
    1-2 μg/ml, 60 min
    Positive control:
    RBL rat basophilic leukemia cell line
    A-431 human epidermoid carcinoma cell line
    RAMOS lymphoma cell line
    U-937 human histiocytic lymphoma cell line
    JURKAT human peripheral blood T cell leukemia cell line
    Negative control:
    HeLa human cervix carcinoma cell line
    Sample preparation:
    Resuspend approx. 50 mil. cells in 1 ml cold lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate (1:1) with non-reducing SDS-PAGE sample buffer.
    Application note: Non-reducing conditions. SDS-PAGE (12% separating gel).
  • Immunohistochemistry (paraffin sections)
    Recommended dilution: 5 μg/ml
    Positive tissue: tonsil B cells
  • Immunocytochemistry
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

  • *Tolar P, Draberova L, Draber P: Protein tyrosine kinase Syk is involved in Thy-1 signaling in rat basophilic leukemia cells. Eur J Immunol. 1997 Dec;27(12):3389-97. [Abstract]
  • *Amoui M, Draberova L, Tolar P, Draber P: Direct interaction of Syk and Lyn protein tyrosine kinases in rat basophilic leukemia cells activated via type I Fc epsilon receptors. Eur J Immunol. 1997 Jan;27(1):321-8. [Abstract]
  • *Ma H, Yankee TM, Hu J, Asai DJ, Harrison ML, Geahlen RL: Visualization of Syk-antigen receptor interactions using green fluorescent protein: differential roles for Syk and Lyn in the regulation of receptor capping and internalization. J Immunol. 2001 Feb 1;166(3):1507-16. [Abstract] [Full Text]
  • *Swann PG, Odom S, Zhou YJ, Szallasi Z, Blumberg PM, Draber P, Rivera J: Requirement for a negative charge at threonine 60 of the FcRgamma for complete activation of Syk. J Biol Chem. 1999 Aug 13;274(33):23068-77. [Abstract] [Full Text]
  • *Kovarova M, Tolar P, Arudchandran R, Draberova L, Rivera J, Draber P: Structure-function analysis of Lyn kinase association with lipid rafts and initiation of early signaling events after Fcepsilon receptor I aggregation. Mol Cell Biol. 2001 Dec;21(24):8318-28. [Abstract] [Full Text]
  • *Manetz TS, Gonzalez-Espinosa C, Arudchandran R, Xirasagar S, Tybulewicz V, Rivera J: Vav1 regulates phospholipase cgamma activation and calcium responses in mast cells. Mol Cell Biol. 2001 Jun;21(11):3763-74. [Abstract] [Full Text]
  • *Parravicini V, Gadina M, Kovarova M, Odom S, Gonzalez-Espinosa C, Furumoto Y, Saitoh S, Samelson LE, O'Shea JJ, Rivera J: Fyn kinase initiates complementary signals required for IgE-dependent mast cell degranulation. Nat Immunol. 2002 Aug;3(8):741-8. Epub 2002 Jul 1. [Abstract]
  • Product Specific References

  • *Arudchandran R, Brown MJ, Peirce MJ, Song JS, Zhang J, Siraganian RP, Blank U, Rivera J: The Src homology 2 domain of Vav is required for its compartmentation to the plasma membrane and activation of c-Jun NH(2)-terminal kinase 1. J Exp Med. 2000 Jan 3;191(1):47-60. [Abstract] [Full Text]
  • *Halova I, Draberova L, Draber P: A novel lipid raft-associated glycoprotein, TEC-21, activates rat basophilic leukemia cells independently of the type 1 Fc epsilon receptor. Int Immunol. 2002 Feb;14(2):213-23. [Abstract] [Full Text]
  • *Heneberg P, Lebduska P, Draberova L, Korb J, Draber P: Topography of plasma membrane microdomains and its consequences for mast cell signaling. Eur J Immunol. 2006 Oct;36(10):2795-806. [Abstract]
  • *Osicka R, Osickova A, Hasan S, Bumba L, Cerny J, Sebo P: Bordetella adenylate cyclase toxin is a unique ligand of the integrin complement receptor 3. Elife. 2015 Dec 9;4:e10766. [Abstract] [Full Text]
  • For research use only. Not for drug, diagnostic or other use.

    Related Products

  • Mouse IgG1 Isotype Control
  • Example Data


    SYK-01
    Fig.1. Western Blotting


    Fig. 1. Western Blotting analysis (non-reducing conditions) of whole cell lysate of RAMOS human Burkitt lymphoma cell line (1), RBL rat basophilic leukemia cell line (2) and HeLa human cervix carcinoma cell line (3, 4).
    Lane 1, 2: immunostaining with anti-Syk (SYK-01)
    Lane 3, 4: immunostaining with anti-human Cytokeratin 18 (DC-10; cat. no. 11-107-C100)
    Lane 5: negative control




    Syk lid fib
    Fig. 2.

    Fig. 2. Immunofluorescence staining of Syk in human primary fibroblasts using anti-Syk (SYK-01; green). Actin cytoskeleton was decorated by phalloidin (red) and cell nuclei stained with DAPI (blue).










    Syk HeLa
    Fig. 3.

    Fig. 3. Immunofluorescence staining of Syk in human HeLa cell line using anti-Syk (SYK-01; green). Actin cytoskeleton was decorated by phalloidin (red) and cell nuclei stained with DAPI (blue).









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