LAT [pY132] Antibody | catalog# XBP-4155
Background
Linker for activation of T cells (LAT) is a 36 kDa transmembrane adapter protein that is tyrosine-phosphorylated following T-cell receptor (TCR) stimulation by ZAP-70 and Syk. Four distal tyrosine residues (Y132, Y171, Y191 and Y226 in human LAT) are crucial for its activity and subsequent signaling to downstream molecules. Tyrosine 132 mediates LAT binding to PLC?-1, and is necessary for TCR-mediated calcium mobilization and activation of ERK and NFAT proteins.
Description
![LAT [pY132] Antibody](./image/product/04/17/XBP-4155.gif)
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 LAT [pY132] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide 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.
Specifications
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Clonality/Clone
This is a polyclonal antibody.Specificity
Human LAT. Mouse (85% homologous) and rat (77%) LAT have not been tested, but are expected to react.Source
LAT antibody was produced against a chemically synthesized phosphopeptide derived from a region of human LAT that contains tyrosine 132.Purification
LAT antibody was 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.Host
LAT [pY132] antibody was raised in rabbit.Please use anti-rabbit secondary antibodies.
Application
For Western blotting, we recommend using the antibody at 0.1-1.0 μg/mL. At 0.35 μg/mL, the dilution provides 100 mL working solution, which at 10 mL/blot allows 10 blots to be performed.Tested Application
WBBuffer
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Storage
Store at -80˚C. Upon initial thawing, apportion into working aliquots and Store at -80˚C. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody.Positive Control
- Jurkat E6.1 cells]
Species Reactivity
H, MReference
- Perez-Villar, J.J., et al. (2002) Phosphorylation of the linker for activation of T-cells by Itk promotes recruitment of Vav. Biochemistry 41(34):10732-10740.
- Sommers, C.L., et al. (2002) A LAT mutation that inhibits T cell development yet induces lymphoproliferation. Science 296(5575):2040-2043.
- Lin, J., and A. Weiss (2001) Identification of the minimal tyrosine residues required for linker for activation of T cell function. J. Biol. Chem. 276(31):29588-29595.
- Zhang, W., et al. (2000) Association of Grb2, Gads, and phospholipase C-gamma 1 with phosphorylated LAT tyrosine residues. Effect of LAT tyrosine mutations on T cell antigen receptor-mediated signaling. J. Biol. Chem. 275(30):23355-23361.]