Biolegend, 933101, Purified anti-Lck Phospho (Tyr394) Antibody, 25μg

The Src family tyrosine kinase p56 Lck (Lck) is a non-receptor tyrosine kinase that plays a critical role in T cell selection and maturation within the thymus, and also in the function of mature T cells. Lck, which is constitutively bound to cytosolic domains of CD4 and CD8 surface receptors, plays an essential role in T cell receptor (TCR) signaling. Engagement of the TCR with peptide antigen-loaded MHC complexes results in the recruitment of CD4- and CD8-bound Lck to the TCR/CD3 signaling complex. Lck then transphosphorylates TCR-gamma chains and CD3 subunits, thereby activating the TCR/CD3 signaling pathway and leading to the recruitment and subsequent phosphorylation of Zap70 by Lck. Lck also plays an important role in interleukin-2 signaling that regulates the T cell proliferative response. Phosphorylation of Lck by CSK at tyrosine 505 negatively regulates the kinase, and is proposed to generate a closed, inactive conformation of the protein. Conversely, phosphorylation at tyrosine 394 in the activation loop of Lck greatly stimulates enzymatic activity. In many types of cancer, Lck is a proliferative and anti-apoptotic driver, and has been proposed as a target for therapeutic intervention.
25μg
Verified Reactivity: Human, Mouse
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Synthetic peptide corresponding to human Src phosphorylated at tyrosine 419
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application: WB - Quality tested ICC, ICFC - Verified
Recommended Usage: Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For intracellular flow cytometry using our True-Phos™ Perm Buffer in Cell Suspensions Protocol, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: This clone may recognize other Src family members. This clone was developed using a synthetic peptide corresponding to human Src phosphorylated at tyrosine 419, which displays high sequence homology with the region surrounding Lck tyrosine 394. To confirm specificity to Lck Phospho (Tyr394), the cell line J.Cam1.6 was used. This cell line is a clonal derivative of Jurkat cells that harbors a truncated LCK allele with defective protein expression (Straus, et al. 1992. Cell. 70:585).
Additional Product Notes: This clone has not been tested for IP. This clone was ICC tested on PFA-fixed cells using both methanol and Triton X-100 permeabilization methods. Both permeabilazation methods were compatible with staining. Triton X-100 permeabilization produces stronger staining. During ICC product development testing, this clone produced strong nuclear staining in a small subpopulation (<1%) of both unstimulated and stimulated cells. The source of this staining is not clear but we do not believe it is Lck-dependent.
Product Citations: Lam W, et al. 2023. Front Pharmacol. 13:1095186. PubMed Sasai M, et al. 2021. J Exp Med. 218:. PubMed
RRID: AB_2814639 (BioLegend Cat. No. 933101) AB_2814640 (BioLegend Cat. No. 933102)
Structure: Lck is a 509 amino acid protein with a predicted molecular weight of 58 kD
Distribution: Plasma-membrane associated/T cells
Function: Protein Tyrosine Kinase
Cell Type: T cells
Biology Area: Cell Biology, Signal Transduction
Molecular Family: Protein Kinases/Phosphatase
Antigen References: Phillipsen L, et al. 2017. Sci Signal. 10:eaaf4736. Moogk D, et al. 2016. J Immunol. 197:644. Klammt C, et al. 2015. Nat Immunol. 16:961. Cancer Genome Atlas Network. 2015. Cell. 161:1681. Casas J, et al. 2014. Nat Commun. 5:5624 McNeill L, et al. 2007. Immunnity. 27:425. Lefebvre DC, et al. 2003. Biochim Biophys Acta. 1650:40. Kabouridis PS. et al. 2003. Biochem J. 371:907. Abraham N, et al. 1990. Mol. Cell. Biol. 10:5197.
Gene ID: 3932
UniProt: View information about Lck Phospho Tyr394 on UniProt.org
Clone: A18002D
Regulatory Status: RUO
Other Names: Lsk, p56lck, LCK Proto-Oncogene, Src Family Tyrosine Kinase, Tyrosine-protein kinase LCK, Leukocyte C-terminal Src kinase, LSK, Lymphocyte cell-specific protein-tyrosine kinase, T cell-specific protein-tyrosine kinase
Isotype: Mouse IgG1, κ