HTRF Mouse Phospho-SLP76 (Ser376) Detection Kit, 500 Assay Points

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HTRF Mouse Phospho-SLP76 (Ser376) Detection Kit, 500 Assay Points

Slp-76 P-s376 (m) Kit - 50,000 Tests

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

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Feature	Specification
Application	Cell Signaling
Sample Volume	16 µL

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Slp-76 P-s376 (m) Kit - 50,000 Tests

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Product Variants

Unit Size: 500 Assay Points

Part #:

63ADK109PEG

List Price

USD 2,147.00

Unit Size: 10,000 Assay Points

Part #:

63ADK109PEH

List Price

USD 12,490.00

Product information

Overview

This HTRF cell-based assay enables rapid, quantitative detection of mouse SLP-76 phosphorylated on Serine 376. Phospho-SLP-76 creates a scaffold on which key signaling complexes are built, and is a marker of T-lymphocyte activation.

Specifications

Other Specifications

Application	Cell Signaling
Brand	HTRF
Detection Modality	HTRF
Lysis Buffer Compatibility	Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 4
Molecular Modification	Phosphorylation
Product Group	Kit
Sample Volume	16 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Phosphoproteins
Target Species	Mouse
Technology	TR-FRET
Therapeutic Area	Oncology & Inflammation
Unit Size	500 Assay Points

Video gallery

How it works

Mouse phospho-SLP-76 (Ser376) assay principle

The Mouse phospho-SLP-76 (Ser376) assay measures mouse SLP-76 when phosphorylated at Ser376. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Mouse phospho-SLP-76 (Ser376) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, and the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the protein’s phosphorylation state under a no-wash assay format.

Mouse phospho-SLP-76 (Ser376) 2-plate assay protocol

The 2 plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the addition of mouse phospho-SLP-76 (Ser376) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

2biomarkers-how-it-works-assay-protocol-alpha-tubulin-63adk072peg-63adk072peh.svg

Mouse phospho-SLP-76 (Ser376) 1-plate assay protocol

Detection of mouse phosphorylated SLP-76 (Ser376) with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.

2phospho-how-it-works-total-btk-1-plate-assay-protocol.svg

Simplified pathway

Function and regulation of SLP-76

TCR engagement promotes the phosphorylation of immunoreceptor tyrosine-based activation motifs (ITAMs) on the cytosolic side of the TCR/CD3 complex by lymphocyte protein tyrosine kinase (Lck). Zap-70 is recruited to the TCR/CD3 complex, where it becomes phosphorylated and activated. ZAP-70 phosphorylates SLP-76. Activated SLP-76 translocates to the plasma membrane and promotes a multi-protein complex, which participates in the activation, survival, and proliferation of T-Lymphocytes.