Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb #14745

No. Size Price
14745S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
14745T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
14745 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 76 Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Specificity / Sensitivity

Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb recognizes endogenous levels of SLP-76 protein only when phosphorylated at Ser376.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser376 of human SLP-76 protein.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with H2O2 (11 mM, 1 min; green), using Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human PBMCs, unstimulated (left) or stimulated (right) with anti-CD3 and anti-CD28 (both at 2 μg/ml, 1 hr, 37ºC), using Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb and co-stained with a CD3 antibody. Stimulation increases phosphorylation of SLP-76 at (Ser376) in the CD3 (T cell) population (right plot, upper right quadrant) relative to unstimulated cells (left plot, upper right quadrant). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from EL4 cells, untreated (-) or treated with H2O2 (11 mM, 1 min; +), using Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb (upper) or SLP-76 Antibody #4958 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with H2O2 (11 mM, 1 min; +), using Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb (upper) or SLP-76 Antibody #4958 (lower).

Background

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 down regulation of TCR signaling (6-8).

  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. Shui, J.W. et al. (2007) Nat Immunol 8, 84-91.
  7. Di Bartolo, V. et al. (2007) J Exp Med 204, 681-91.
  8. Lasserre, R. et al. (2011) J Cell Biol 195, 839-53.

Application References

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

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Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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