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Phospho-LAT (Tyr220) (E3S5L) Rabbit mAb #20172
Filter:
- WB
- IP
Western blot analysis of extracts from Jurkat and EL4 cells, treated with isotype control (-) or treated with either cross-linked anti-CD3 plus anti-CD28 (10 μg/mL each, 5 min; +) or H2O2 (11 mM, 1 min; +), using Phospho-LAT (Tyr220) (E3S5L) Rabbit mAb (upper), LAT Antibody #9166 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Supporting Data
| REACTIVITY | H M |
| SENSITIVITY | Endogenous |
| MW (kDa) | 40 |
| Source/Isotype | Rabbit IgG |
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 | 1:200 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #84593.
For a carrier free (BSA and azide free) version of this product see product #84593.
Protocol
Specificity / Sensitivity
Phospho-LAT (Tyr220) (E3S5L) Rabbit mAb recognizes endogenous levels of LAT protein when phosphorylated at Tyr220. This antibody shows cross-reactivity with phosphorylated EGFR, and detects a 70 kDa band of unknown origin in some treated cell lines. Tyr220 of LAT, long isoform corresponds to Tyr191 of LAT, short isoform. Weak cross-reactivity with phosphorylated tyrosines 171 and 255 of LAT has been observed.
Species Reactivity:
Human, Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr220 of human LAT protein.
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).
Pathways
Explore pathways related to this product.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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