Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated LAT. The final product is generated by
affinity chromatography using an LAT-derived peptide that is phosphorylated at tyrosine 132.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 31.4 kDa).
Required for TCR (T-cell antigen receptor)- and pre-TCR-mediated signaling, both in mature T-cells and during their development. Involved in FCGR3 (low affinity immunoglobulin gamma Fc region receptor III)-mediated signaling in natural killer cells and FCER1 (high affinity immunoglobulin epsilon receptor)-mediated signaling in mast cells. Couples activation of these receptors and their associated kinases with distal intracellular events such as mobilization of intracellular calcium stores, PKC activation, MAPK activation or cytoskeletal reorganization through the recruitment of PLCG1, GRB2, GRAP2, and other signaling molecules.
Expressed in thymus, T-cells, NK cells, mast cells and, at lower levels, in spleen. Present in T-cells but not B-cells (at protein level).
Phosphorylated on tyrosines by ZAP-70 upon TCR activation, or by SYK upon other immunoreceptor activation; which leads to the recruitment of multiple signaling molecules. Is one of the most prominently tyrosine-phosphorylated proteins detected following TCR engagement. May be dephosphorylated by PTPRJ. Palmitoylation of Cys-26 and Cys-29 is required for raft targeting and efficient phosphorylation.
Linker for activation of T cells family member 1 antibody
linker for activation of T cells, transmembrane adaptor antibody
Linker for activation of T-cells family member 1 antibody
p36 38 antibody
Western blot - LAT (phospho Y132) antibody (ab4476)
Predicted band size : 31.4 kDa
Extracts prepared from Jurkat E6.1 cells were left unstimulated (1) or stimulated (2-5), resolved by SDS-PAGE on a 10% polyacrylamide gel, and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.35 µg/ml phospho LAT (Tyr 132) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the nonphosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to phospho LAT (Tyr 132) completely blocks the antibody signal, thereby demonstrating the specificity of the antibody, and stimulation-induced tyrosine phosphorylation of phospho LAT (Tyr 132).