WB, W-S, IP, IHC-Bond, IHC-P, FC-FP
H M
Endogenous
76
Rabbit IgG
#Q13094
3937
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:50 - 1:250 |
| Immunoprecipitation | 1:50 |
| IHC Leica Bond | 1:200 - 1:800 |
| Immunohistochemistry (Paraffin) | 1:200 - 1:800 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 - 1:400 |
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 #10190.
Specificity / Sensitivity
Species Reactivity:
Human, Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser376 of human SLP-76 protein.
Background
SH2 domain-containing leukocyte protein of 76 kDa (SLP-76) is a hematopoietic adaptor protein that is important in multiple biochemical signaling pathways and necessary for T cell development and activation (1). ZAP-70 phosphorylates SLP-76 and LAT as a result of TCR ligation. SLP-76 has amino-terminal tyrosine residues followed by a proline-rich domain and a carboxy-terminal SH2 domain. Phosphorylation of Tyr113 and Tyr128 result in recruitment of the GEF Vav and the adaptor protein Nck (2). TCR ligation also leads to phosphorylation of Tyr145, which mediates an association between SLP-76 and Itk, which is accomplished in part via the proline-rich domain of SLP-76 and the SH3 domain of Itk (3). Furthermore, the proline-rich domain of SLP-76 binds to the SH3 domains of Grb2-like adaptor Gads (3,4). In resting cells, SLP-76 is predominantly in the cytosol. Upon TCR ligation, SLP-76 translocates to the plasma membrane and promotes the assembly of a multi-protein signaling complex that includes Vav, Nck, Itk, and PLCγ1 (1). The expression of SLP-76 is tightly regulated; the protein is detected at very early stages of thymocyte development, increases as thymocyte maturation progresses, and is reduced as cells mature to CD4+ CD8+ double-positive thymocytes (5).
Following TCR ligation, SLP-76 is phosphorylated at Ser376 by the hematopoietic progenitor kinase 1 (HPK1) (6,7). This phosphorylation induces interaction with 14-3-3ε, which leads to the disassembly of TCR signaling complexes and downregulation of TCR signaling (6-8).
- Clements, J.L. (2003) Immunol Rev 191, 211-9.
- Bubeck Wardenburg, J. et al. (1998) Immunity 9, 607-16.
- Bunnell, S.C. et al. (2000) J Biol Chem 275, 2219-30.
- Liu, S.K. et al. (1999) Curr Biol 9, 67-75.
- Clements, J.L. et al. (1998) J Immunol 161, 3880-9.
- Shui, J.W. et al. (2007) Nat Immunol 8, 84-91.
- Di Bartolo, V. et al. (2007) J Exp Med 204, 681-91.
- Lasserre, R. et al. (2011) J Cell Biol 195, 839-53.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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