Lck [pY505] Antibody | catalog# XBP-4160
Background
Lck (p56lck), a member of the Src family of non-receptor tyrosine protein kinases, is expressed predominantly in T-cells. Lck function is critical for both T-cell development in the thymus and activation of mature T-cells in the periphery by antigen. The activity of Lck is regulated by phosphorylation of multiple sites, primarily the two conserved residues, tyrosines 394 (equivalent to 418 in Src) and 505 (equivalent to 529 in Src). When phosphorylated, carboxy-terminal Tyr-505 associates intramolecularly with the SH2 domain in the amino-terminal half of the protein, stabilizing Lck in a conformation that, biologically, is relatively inactive. In the absence of phosphorylation at Tyr-505, intramolecular binding of the carboxyl terminus to the SH2 domain does not occur, and Lck exhibits increased activity in vivo.
Description
![Lck [pY505] Antibody](./image/product/11/20/XBP-4160.gif)
Full length untagged recombinant human Lck protein added to background cell extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, and then incubated with the Lck [pY505] antibody in a 1% BSA-TBST buffer for two hours at room temperature, following prior incubation with: the phosphopeptide immunogen (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), the non-phosphopeptide derived from the corresponding region of Src (3), a generic phosphotyrosine-containing peptide (4), no peptide (5) or the phosphopeptide derived from the corresponding region of Src (6). After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase conjugate and signals were detected using the Tropix WesternStar™ detection method.
Specifications
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Clonality/Clone
This is a polyclonal antibody.Specificity
Human Lck. Mouse Lck [pY505] (100% homology), chicken Lck [pY505] (91.7% homology), Hck [pY522] (75% homology) and Lyn [pY508] (83.3% homology) have not been tested.Source
Lck antibody was produced against a chemically synthesized phosphopeptide derived from the region of human Lck that contains tyrosine 505 (based on Swiss Protein database, accession number P06239). The sequence is conserved in mouse.Purification
Lck antibody was purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Lck, and ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphoserine (irrespective of the sequence). The final product is generated by affinity chromatography using a Lck-derived peptide that is phosphorylated at tyrosine 505.Host
Lck [pY505] antibody was raised in rabbit.Please use anti-rabbit secondary antibodies.
Application
For Western blotting applications, we recommend using the antibody at a 1:1000 starting dilution.Tested Application
WBBuffer
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Storage
Store at -20˚C. We recommend a brief centrifugation before opening to settle vial contents. Then, apportion into working aliquots and store at -20˚C. For shipment or short-term storage (up to one week), 2 - 8˚C is sufficient.Positive Control
- Full length untagged recombinant human Lck protein.]
Species Reactivity
H, M, CReference
- Budagian, V., et al. (2003) Signaling through P2X7 receptor in human T cells involves p56(lck), kinases, and transcription factors AP-1 and NF-kappa B. J. Biol. Chem. 278(3):1549-1560.
- Mustelin, T. and K. Tasken (2003) Positive and negative regulation of T-cell activation through kinases and phosphatases. Biochem. J. 371(Pt 1):15-27.
- Goldmann, W.H. (2002) p56(lck) Controls phosphorylation of filamin (ABP-280) and regulates focal adhesion kinase (pp125(FAK)). Cell Biol. Int. 26(6):567-571.
- Gervais, F.G. and A. Veillette (1997) Reconstitution of interactions between protein-tyrosine phosphatase CD45 and tyrosine-protein kinase p56(lck) in nonlymphoid cells. J. Biol. Chem. 272(19):12754-12761.]