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Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (Alexa Fluor^® 647 Conjugate)

R Recombinant

Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (Alexa Fluor^® 647 Conjugate) #40887

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Supporting Data

REACTIVITY	H
SENSITIVITY	Endogenous
MW (kDa)
Source/Isotype	Rabbit IgG

Application Key:

F-Flow Cytometry

Species Cross-Reactivity Key:

H-Human

Product Information

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor^® 647 fluorescent dye and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-SLP-76 (Ser376) (D7S1K) XP^® Rabbit mAb #92711.

Product Usage Information

Application	Dilution
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

Specificity / Sensitivity

Phospho-SLP-76 (Ser376) (D7S1K) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate) recognizes endogenous levels of SLP-76 protein only when phosphorylated at Ser376.
Clone E3G9U is more sensitive by flow cytometry than clone D7S1K.

Species Reactivity:

Human

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).

Database Links

UniProt ID: Q13094

Entrez-Gene Id: 3937

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Alexa Fluor is a registered trademark of Life Technologies Corporation.

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Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (Alexa Fluor® 647 Conjugate) #40887