Thermo Fisher Scientific Phospho-SLP-76 (Tyr128) Monoclonal Antibody (HNDZ55), APC, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.5 µg)/test

-

Product Specifications

Species Reactivity

Human

Host/Isotype

Mouse / IgG2b, kappa

Recommended Isotype Control

Mouse IgG2b kappa Isotype Control (eBMG2b), APC, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

HNDZ55

Conjugate

APC APC APC

Excitation/Emission Max

651/660 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2573284

Product Specific Information

Description: This HNDZ55 monoclonal antibody recognizes human SH2 domain-containing leukocyte protein of 76 kD (SLP-76) when phosphorylated on tyrosine 128 (Y128). SLP-76 is phosphorylated by ZAP-70 downstream of the T cell receptor (TCR). SLP-76 contains an acidic region that includes a PEST domain, several tyrosine residues that are phosphorylated following TCR ligation, a proline-rich domain, and an SH2 domain. Numerous proteins associate with SLP-76 including Vav1, GADS, and PLC gamma 1, supporting the notion that SLP-76 functions as a critical adaptor protein downstream of the TCR. SLP-76-deficient T cell lines or mice deficient in SLP-76 have shown that SLP-76 plays a positive role in promoting T cell development and activation as well as mast cell and platelet function.

This antibody recognizes only human SLP-76 when phosphorylated on Y128. It is not recommended for staining of mouse cells.

Applications Reported:This HNDZ55 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This HNDZ55 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Staining Protocol: Protocol A and Protocol C are recommended for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to Clone Performance Following Fixation/Permeabilization located in the BestProtocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: Staining Intracellular Antigens for Flow Cytometry Protocol located in the BestProtocols® Section under the Resources tab online.

Excitation: 633-647 nm; Emission: 660 nm; Laser: Red Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

SLP-76 was originally identified as a substrate of the ZAP-70 protein tyrosine kinase following T cell receptor (TCR) ligation in the leukemic T cell line Jurkat. The SLP-76 locus has been localized to human chromosome 5q33 and the gene structure has been partially characterized in mice. The human and murine cDNAs both encode 533 amino acid proteins that are 72% identical and comprised of three modular domains. The NH2-terminus contains an acidic region that includes a PEST domain and several tyrosine residues which are phosphorylated following TCR ligation. SLP-76 also contains a central proline-rich domain and a COOH-terminal SH2 domain. A number of additional proteins have been identified that associate with SLP-76 both constitutively and inducibly following receptor ligation, supporting the notion that SLP-76 functions as an adaptor or scaffold protein. Studies using SLP-76 deficient T cell lines or mice have provided strong evidence that SLP-76 plays a positive role in promoting T cell development and activation as well as mast cell and platelet function.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.