Phospho-LAT (Tyr255) Antibody #45170
Filter:
- WB
- IP
Western blot analysis of extracts from serum-starved Jurkat and EL4 cells, treated with isotype control (-) or treated with cross-linked anti-CD3 plus anti-CD28 (+; 10 ug/mL each, 5 min), using Phospho-LAT (Tyr255) Antibody (upper) and LAT Antibody #9166 (lower).
Supporting Data
| REACTIVITY | H M |
| SENSITIVITY | Endogenous |
| MW (kDa) | 40 |
| SOURCE | Rabbit |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- Related Products
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Protocol
Specificity / Sensitivity
Phospho-LAT (Tyr255) Antibody recognizes endogenous levels of LAT protein only when phosphorylated at Tyr255. Tyr255 of LAT, long isoform corresponds to Tyr226 of LAT, short isoform.
Species Reactivity:
Human, Mouse
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr255 of human LAT protein, isoform 1. Antibodies are purified by protein A and peptide affinity chromatography.
Background
LAT, a transmembrane adaptor protein expressed in T, NK, and mast cells, is an important mediator for T cell receptor (TCR) signaling (1). Upon TCR engagement, activated Zap-70 phosphorylates LAT at multiple conserved tyrosine residues within SH2 binding motifs, exposing these motifs as the docking sites for downstream signaling targets (2,3). The phosphorylation of LAT at Tyr171 and Tyr220 enables the binding of Grb2, Gads/SLP-76, PLCγ1, and PI3 kinase through their SH2 domain and translocates them to the membrane. This process eventually leads to activation of the corresponding signaling pathways (1-4).
Research studies have shown that LAT is also phosphorylated at Tyr225 by ZAP-70 and Syk following activation of the TCR, which facilitates its binding to Grb2 and Vav and subsequent downstream activation of ERK (5-7).
Research studies have shown that LAT is also phosphorylated at Tyr225 by ZAP-70 and Syk following activation of the TCR, which facilitates its binding to Grb2 and Vav and subsequent downstream activation of ERK (5-7).
- Wonerow, P. and Watson, S.P. (2001) Oncogene 20, 6273-83.
- Zhang, W. et al. (1998) Cell 92, 83-92.
- Paz, P. E. et al. (2001) Biochem. J. 356, 461-71.
- Zhang, W. et al. (2000) J. Biol. Chem. 275, 23355-61.
- Zhang, W. et al. (2000) J Biol Chem 275, 23355-61.
- Paz, P.E. et al. (2001) Biochem J 356, 461-71.
- Lin, J. and Weiss, A. (2001) J Biol Chem 276, 29588-95.
Pathways
Explore pathways related to this product.
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