CST, 15642S, Phospho-SLP-76 (Ser376) (E3G9U) Rabbit Monoclonal Antibody (Pacific Blue™ Conjugate)

Monoclonal Antibody for studying SLP76 (Ser376) phosphate. Validated for Flow Cytometry (Fixed/Permeabilized). Highly specific and rigorously validated in-house, Phospho-SLP-76 (Ser376) (E3G9U) Rabbit Monoclonal Antibody (Pacific Blue^™ Conjugate) (CST #15642) is ready to ship. Product Usage Information Flow Cytometry (Fixed/Permeabilized): 1:50 Storage Supplied in PBS (pH 7.2), less than 0.1% sodium azide, and 2 mg/mL BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze. Protocol Available protocols: Flow Cytometry (Fixed/Permeabilized) Specificity / Sensitivity Phospho-SLP-76 (Ser376) (E3G9U) Rabbit Monoclonal Antibody (Pacific Blue™ Conjugate) recognizes endogenous levels of SLP-76 protein only when phosphorylated at Ser376. Non-specific staining was observed in human stomach crypts by immunohistochemistry. Clone E3G9U is more sensitive by flow cytometry than clone D7S1K. Species Reactivity: Human, Mouse Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide 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). Alternate Names 76 kDa tyrosine phosphoprotein; LCP2; Lymphocyte cytosolic protein 2; lymphocyte cytosolic protein 2 (SH2 domain containing leukocyte protein of 76kDa); SH2 domain-containing leukocyte protein of 76 kDa; SH2 domain-containing leukocyte protein of 76kD; SLP-76; SLP-76 tyrosine phosphoprotein; SLP76 Specification REACTIVITY: H M SENSITIVITY: Endogenous Source/Isotype: Rabbit IgG