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FastScan^™ Phospho-SLP-76 (Ser376) ELISA Kit

FastScan^™ Phospho-SLP-76 (Ser376) ELISA Kit #25424

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ELISA

Supporting Data

REACTIVITY

Application Key:

ELISA-ELISA

Species Cross-Reactivity Key:

H-Human

Product Information

Product Description

The FastScan™ Phospho-SLP-76 (Ser376) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of SLP-76 when phosphorylated at Ser376. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with phospho-SLP-76 (Ser376) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of phospho-SLP-76 (Ser376).

*Antibodies in this kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

The FastScan™ Phospho-SLP-76 (Ser376) ELISA Kit detects endogenous levels of SLP-76 when phosphorylated at Ser376 as shown in Figure 1. This kit is able to detect phospho-SLP-76 (Ser376) in human samples but is not cross-reactive with mouse phospho-SLP-76 (Ser376), as determined through in-house testing, but may also detect homologous proteins from other species. The FastScan™ Phospho-SLP-76 (Ser376) ELISA Kit #25424 provides better performance when detecting human phospho-SLP-76 (Ser376) as compared to kit #30794.

Species Reactivity:

Human

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).

Database Links

UniProt ID: Q13094

Entrez-Gene Id: 3937

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

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

FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.

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