Description
The non-receptor protein tyrosine kinases of the Src family have similar domain structures with considerable sequence homologies. They are associated with cell membranes and are activated by a wide variety of cell-surface receptors. Through their participation in cell-signaling cascades that mediate an array of cellular responses, they control essential cellular activities. Src, the prototype of the Src family, is encoded by the SRC protooncogene, which is a homolog of the v-Src gene of the Rous sarcoma virus. Src's oncogenic potential may be a consequence of its role as a regulator of cell growth, morphology, motility, and adhesion. The protein contains domains that mediate protein-protein interactions and regulation of its tyrosine kinase activity. Mechanisms that regulate Src activity include phosphorylations of serines and tyrosines in most of the domains and interactions with receptor proteins, integrins, and other binding proteins. In particular, autophosphorylation at tyrosine 418 (Y418) activates tyrosine kinase activity. Phosphorylations at other sites are mediated by kinases such as cdc2, CSK, and CHK in signaling cascades and may up- or down-regulate Src activity. The K98-37 monoclonal antibody recognizes the phosphorylated Y418 in the protein kinase domain of activated Src. The orthologous phosphorylation sites in chicken, mouse, and rat Src are Y415, Y423, and Y418, respectively. Because of sequence similarities in this region, it may also recognize the homologous sites of some other Src family members: Lyn (pY396), Fyn (pY420), Hck (pY410), Lck (pY394), and Yes (pY425).
Either BD Cytofix™ fixation buffer or BD Phosflow™ Fix Buffer I may be used for cell fixation. 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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. 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.
4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
6. 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.
7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
