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Analysis of SLP-76 in human peripheral blood lymphocytes. Human whole blood was lysed and fixed with 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10-15 minutes at 37ºC, permeabilized (BD Phosflow™ Perm Buffer II, Cat. No. 558052) on ice for 30 minutes, stained with Alexa Fluor® 647 Mouse Anti-Human CD3 mAb UCHT1 (Cat. No. 557706), PerCP-Cy5.5 anti-human CD20 mAb H1(FB1) (Cat. No. 558021), and either PE Mouse anti-SLP-76 (Cat. No. 560058, solid-line histograms) or PE Mouse IgG2a, κ Isotype control mAb MOPC-173 (Cat. No. 558595, dashed-line histograms). The figures show lymphocyte subpopulations that were selected by their scatter profile and surface antigen expression. SLP-76 expression on CD3-positive CD20-negative T lymphocytes (left panel), CD20-positive CD3-negative B lymphocytes (center panel), and CD20-negative CD3-negative NK cells (right panel) are displayed. There was no detectable expression of SLP-76 in the monocyte/granulocyte population (not shown). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
BD™ Phosflow PE Mouse anti-SLP-76
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
SLP-76 (SH2 domain-containing Leukocyte Protein of 76 kDa) is a tyrosine phosphoprotein that is involved in the T cell receptor (TCR)-mediated intracellular signaling pathway. It may be involved in the signaling pathways of other peripheral blood leukocytes; thymic/splenic cells; and in human T, B, and monocytic cell lines. SLP-76 consists of several motifs that signify its importance in protein-protein interactions involved in intracellular signaling pathways, such as the SH2 domain in the C-terminus, the three amino-terminus 17-amino acid repeats with conserved tyrosine and acidic residues (DYE(S/P)P), and a proline rich region. SLP-76 has been shown to associate with Gads, Grb2, PLCγ1, SLAP-130, and Vav, all of which are part of the signaling cascade in T lymphocytes. An early event in the T cell activation pathway is the phosphorylation, by the Syk-family kinase ZAP-70, of SLP-76 at the three conserved tyrosine motifs, which then mediate interactions with downstream effectors.
The H3 monoclonal antibody recognizes SLP-76, regardless of phosphorylation status.
Development References (5)
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Bonilla FA, Fujita RM, Pivniouk VI, Chan AC, Geha RS. Adapter proteins SLP-76 and BLNK both are expressed by murine macrophages and are linked to signaling via Fcγ receptors I and II/III. Proc Natl Acad Sci U S A. 2000; 97(4):1725-1730. (Clone-specific: Immunoprecipitation).
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Fang N, Motto DG, Ross SE, Koretzky GA. Tyrosines 113, 128, and 145 of SLP-76 are required for optimal augmentation of NFAT promoter activity. J Immunol. 1996; 157:3769-3773. (Biology). View Reference
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Janssen E, Zhang W. Adaptor proteins in lymphocyte activation. Curr Opin Immunol. 2003; 15:269-276. (Biology). View Reference
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Wardenburg JB, Fu C, Jackman JK, et al. Phosphorylation of SLP-76 by the ZAP-70 protein-tyrosine kinase is required for T-cell receptor function. J Biol Chem. 1996; 271(33):19641-19644. (Immunogen: Immunoprecipitation, Western blot).
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Wu JN, Koretzky GA. The SLP-76 family of adapter proteins. Semin Immunol. 2004; 16:379-393. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.