Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Antibody Pair #7937

Kit Includes Volume Cap Color
LAT Capture Ab (100X) 0.4 ml Pink
P-LAT (Y191) Detection Ab (100X) 0.4 ml Blue
Anti-rabbit IgG, HRP-linked Antibody 0.04 ml Red

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Important Ordering Details

Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Protocols

Description

CST's PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Kit #7936. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The LAT mouse capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysate is added followed by a phospho-LAT (Tyr191) rabbit detection antibody and HRP-conjugated, anti-rabbit IgG antibody. HRP substrate (TMB) is then added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-LAT (Tyr191).Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Sandwich ELISA

Sandwich ELISA

Figure 1. The relationship between the protein concentration of the lysate from untreated and anti-CD3-treated Jurkat cells and the absorbance at 450 nm using the PathScan® Phospho-LAT (Tyr191) Sandwich ELISA Antibody Pair #7937 is shown. After a 48 hour starvation, Jurkat cells were treated with anti-CD3 (10 µg/ml) for 2 minutes at 37ºC and then lysed.

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 Tyr191 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).

  1. Wonerow, P. and Watson, S.P. (2001) Oncogene 20, 6273-6283.
  2. Zhang, W. et al. (1998) Cell 92, 83-92.
  3. Paz, P. E. et al. (2001) Biochem. J. 356, 461-471.
  4. Zhang, W. et al. (2000) J. Biol. Chem. 275, 23355-23361.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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