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Peptide Competition. Background extracts alone (1) or with added recombinant active human Lck protein (2-5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, and then incubated with the Lck [pS192] antibody in a 1% BSA-TBST buffer for two hours at room temperature, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphotyrosine-containing peptide (4) or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab"e;)2 anti-rabbit IgG HRP conjugate (Cat. no. ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to Lck [pY192] blocks the antibody signal, demonstrating the specificity of the antibody.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the regionof human Lck that contains tyrosine 192 (based on Swiss Protein database, accession number P06239).The sequence is conserved in mouse.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
LCK is a member of the Src family of protein tyrosine kinases (PTKs). This protein is a key signaling molecule in the selection and maturation of developing T-cells. It 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. The protein localizes to the plasma membrane and pericentrosomal vesicles, and binds to cell surface receptors, including CD4 and CD8, and other signaling molecules.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
EC 220.127.116.11; kinase Lck; LCK; leukocyte C-terminal Src kinase; LSK; LSK-T; lymphocyte cell-specific protein-tyrosine kinase; p56(LSTRA) protein-tyrosine kinase; P56-LCK; proto-oncogene tyrosine-protein kinase LCK; t cell-specific protein-tyrosine kinase; T-lymphocyte specific protein tyrosine kinase p56lck
IMD22; LCK; LSK; p56lck; pp58lck; YT16