Description
Syk is a non-receptor protein-tyrosine kinase that is closely related to ZAP70 and plays crucial roles in the development and receptor-mediated signaling of most leukocytes and in vascular integrity. Syk is expressed in hematopoietic cells, including B lymphocytes, immature (CD4, CD8 double-negative and double-positive) thymocytes, and myeloid cells, epithelial cell lines, and normal breast tissue. Mature (CD4 or CD8 single-positive) thymocytes and peripheral αβ TCR-bearing T lymphocytes have very low or undetectable levels of Syk. Syk contributes to the signal transduction process by binding to ITAMs (Immunoreceptor Tyrosine-based Activation Motifs) of immune receptors, including Igα and Igβ (CD79a and b), TCRζ, CD3ε, and FcRγ. Upon receptor activation, Syk binds to phosphorylated ITAMs via its two N-terminal SH2 domains thereby activating Syk and causing tyrosines in the interdomain, between the SH2 and Kinase domains of Syk, to undergo auto-phosphorylation and phosphorylation by Lyn. The tyrosine 348 phosphorylation site (pY348) in human Syk is orthologous to tyrosine 342 in mouse and rat Syk and tyrosine 315 in human ZAP70. This phosphorylated site can act as a binding site for other signaling molecules, such as PLCγ, Vav, and Fgr. The I120-772 antibody is specific for human Syk (pY348) and does not cross-react with phosphorylated Zap70. The orthologous phosphorylation site in mouse and rat Syk is Y342.
This antibody conjugate is suitable for intracellular staining of human lymphoid cell lines, peripheral blood mononuclear cells, and mouse splenocytes using BD Phosflow™ Fix Buffer I or Lyse/Fix Buffer. Any of the three BD Phosflow™ permeabilization buffers may be used.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
4. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
7. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
