Anti-Phospho-LAT (Tyr220) Rabbit Antibody [L15N18]

Catalog No.: F3064

Application: Reactivity: Human, Mouse

Usage Information

Dilution
WB1:1000 IP1:200

Application
WB, IP

Reactivity
Human, Mouse

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
40 kDa

Positive Control	Jurkat; EL4
Negative Control	J.CaM2.5

Datasheet & SDS

Biological Description

Specificity
Phospho-LAT (Tyr220) Rabbit mAb detects endogenous levels of LAT protein when phosphorylated at Tyr220.

Uniprot ID
O43561

Clone
L15N18

Synonym(s)
Linker for activation of T-cells family member 1, 36 kDa phosphotyrosine adapter protein (pp36), p36-38, LAT

Background
Phospho- Linker for activation of T cells (LAT) (Tyr220) is a 233–amino acid, type III transmembrane adaptor protein expressed mainly in T cells, NK cells, and mast cells, where it plays a pivotal role in T cell receptor (TCR) signaling. Structurally, LAT has a short extracellular segment, a single transmembrane helix, and a long cytosolic tail containing multiple conserved tyrosines and two palmitoylated cysteines essential for membrane localization. Upon TCR engagement, ZAP-70 phosphorylates key tyrosines—including Tyr220—creating SH2-binding sites for signaling molecules such as Grb2, Gads/SLP-76, PLCγ1, and PI3K, thereby assembling multiprotein complexes that propagate calcium flux, MAPK activation, cytoskeletal reorganization, and gene transcription. Phosphorylation at Tyr220 is critical for recruiting specific adaptors and enzymes, enabling full activation of downstream pathways, making phospho-LAT (Tyr220) a central node in T cell activation and an important marker of proximal TCR signal transduction.

Background

Phospho- Linker for activation of T cells (LAT) (Tyr220) is a 233–amino acid, type III transmembrane adaptor protein expressed mainly in T cells, NK cells, and mast cells, where it plays a pivotal role in T cell receptor (TCR) signaling. Structurally, LAT has a short extracellular segment, a single transmembrane helix, and a long cytosolic tail containing multiple conserved tyrosines and two palmitoylated cysteines essential for membrane localization. Upon TCR engagement, ZAP-70 phosphorylates key tyrosines—including Tyr220—creating SH2-binding sites for signaling molecules such as Grb2, Gads/SLP-76, PLCγ1, and PI3K, thereby assembling multiprotein complexes that propagate calcium flux, MAPK activation, cytoskeletal reorganization, and gene transcription. Phosphorylation at Tyr220 is critical for recruiting specific adaptors and enzymes, enabling full activation of downstream pathways, making phospho-LAT (Tyr220) a central node in T cell activation and an important marker of proximal TCR signal transduction.

References
https://pubmed.ncbi.nlm.nih.gov/20610546/ https://pubmed.ncbi.nlm.nih.gov/11607829/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%