Phospho-LCK (Tyr505), eFluor 660, clone: SRRCHA, eBioscience™
Mouse Monoclonal Antibody
Manufacturer: Life Technologies LS50907642
p56lck is a membrane-associated non-receptor protein tyrosine kinase that belongs to the Src family of kinases. The 3A5 MAb shows no cross-reaction with other members of the Src protein kinase family. The unique N-terminal domain of p56lck interacts with the cytoplasmic tails of the CD4 and CD8 cell surface glycoproteins. Activated p56lck kinase can increase responsiveness of some T cell hybridomas to antigen. The phosphorylation status and the activity of p56lck is regulated by the CD45 tyrosine protein phosphatase. p56lck plays a critical role in T cell development and activation. In humans, p56lck gene is localized to a site in the genome which undergoes frequent chromosomal abnormalities in lymphomas and neuroblastomas.eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered. LCK is a member of the Src-family tyrosine kinase, which are essential for signaling through the T-cell receptor (TCR) complex. Upon TCR triggering, LCK phosphorylates the ITAM motives in its zeta subunits establishing binding sites for the SH2 domains of the tyrosine kinase ZAP70, which is also phosphorylated by LCK and activated to generate subsequent signaling platforms by phosphorylation of adaptor LAT. The majority of LCK is localized to the plasma membrane, however, there is also a significant fraction associated with the Golgi apparatus which may contribute to Raf activation under conditions of weak stimulation through the TCR. LCK contains N-terminal sites for myristylation and palmitylation, a PTK domain, and SH2 and SH3 domains which are involved in mediating protein-protein interactions with phosphotyrosine-containing and proline-rich motifs, respectively. LCK is also involved in the regulation of apoptosis induced by various stimuli, but not by the death receptors. Diseases associated with LCK include Immunodeficiency 22 and CD45 deficiency. Alternatively splice variants of the LCK gene encoding different isoforms of the protein have been found.
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
|4° C, store in dark, DO NOT FREEZE!|
For Research Use Only.