| 货号 | 92711S |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human, Mouse |
| 应用 | WB,F |
| 目标/特异性 | Phopho-SLP-76 (Ser376) (D7S1K) XP® Rabbit mAb recognizes endogenous levels of SLP-76 protein only when phosphorylated at Ser376. |
| 使用方法 | Western Blotting (1:1000) Flow Cytometry (1:400) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | SH2 domain-containing leukocyte protein of 76 kDa (SLP-76) is a hematopoietic adapter 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 adapter 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 adapter 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). |
| 存放说明 | -20C |
| 计算分子量 | 76 |
| 参考文献 | 1 . Clements, J.L. (2003) Immunol Rev 191, 211-9. 2 . Bubeck Wardenburg, J. et al. (1998) Immunity 9, 607-16. 3 . Bunnell, S.C. et al. (2000) J Biol Chem 275, 2219-30. 4 . Liu, S.K. et al. (1999) Curr Biol 9, 67-75. 5 . Clements, J.L. et al. (1998) J Immunol 161, 3880-9. 6 . Di Bartolo, V. et al. (2007) J Exp Med 204, 681-91. 7 . Shui, J.W. et al. (2007) Nat Immunol 8, 84-91. 8 . Lasserre, R. et al. (2011) J Cell Biol 195, 839-53. |
Western blot analysis of extracts from serum-starved Jurkat cells, untreated (-) or treated (+) with either H2O2 (11 mM, 1 min) or cross-linked anti-CD3 plus anti-CD28 (10 μg/mL each, 15 min), using Phospho-SLP-76 (Ser376) (D7S1K) XP® Rabbit mAb (upper), SLP-76 (D1R1A) Rabbit mAb #70896 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with calf intestinal phosphatase (CIP) plus lambda phosphatase (+), using Phospho-SLP-76 (Ser376) (D7S1K) XP® Rabbit mAb (upper), SLP-76 (D1R1A) Rabbit mAb #70896 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left column) or treated with cross-linked anti-CD3 plus anti-CD28 (10 μg/ml each, 15 min; right column), using Phospho-SLP-76 (Ser376) (D7S1K) XP® Rabbit mAb (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (bottom row), and co-stained with a CD3 antibody. Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. |
