|Detection of Human Phospho-LAT (Y161) by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with Pervanadate (PV) for 5 minutes. PVDF Membrane was probed with 0.1 µg/mL of Mouse Anti-Human Phospho-LAT (Y161) Monoclonal Antibody (Catalog # MAB6334) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-LAT (Y161) at approximately 40 kDa (as indicated). For additional reference, the membrane was stripped and reprobed with 1 µg/mL of Sheep Anti-Human LAT Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # AF6208). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
LAT (Linker for Activation of T cells) is a type III transmembrane lipid raft protein that serves as a scaffold for signaling molecules. Isoforms of 262 (36 kDa) and 233, called long and short, respectively, differ by inclusion/exclusion of aa 114‑142; a 269 aa isoform lacks this region, but includes alternate N-terminal sequence. Upon T-cell antigen receptor activation, LAT is multiply phosphorylated by ZAP-70/Syk protein tyrosine kinases, creating docking sites for SH2 domain-containing proteins. Phospholipase C-gamma docks at Y161, which is pY132 in the short form of LAT. Mutation of this site results in a Th2 autoimmune lymphoproliferative disorder in mice.
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