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Alexa Fluor® 647 Mouse anti-SLP-76
Alexa Fluor® 647 Mouse anti-SLP-76

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, then permeabilized (BD Phosflow™ Perm Buffer II, Cat. No. 558052) on ice for 30 minutes, and then stained with Alexa Fluor® 488 Mouse Anti-Human CD3 mAb UCHT1 (Cat. No. 557694), PerCP-Cy5.5 anti-human CD20 mAb H1(FB1) (Cat. No. 558021), and either Alexa Fluor® 647 Mouse anti-SLP-76 (Cat. No. 560057, solid-line histograms) or Alexa Fluor® 647 Mouse IgG2a, κ Isotype control mAb MOPC-173 (Cat. No. 558053, 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.

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, then permeabilized (BD Phosflow™ Perm Buffer II, Cat. No. 558052) on ice for 30 minutes, and then stained with Alexa Fluor® 488 Mouse Anti-Human CD3 mAb UCHT1 (Cat. No. 557694), PerCP-Cy5.5 anti-human CD20 mAb H1(FB1) (Cat. No. 558021), and either Alexa Fluor® 647 Mouse anti-SLP-76 (Cat. No. 560057, solid-line histograms) or Alexa Fluor® 647 Mouse IgG2a, κ Isotype control mAb MOPC-173 (Cat. No. 558053, 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.

Product Details
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BD Phosflow™
Human (QC Testing), Mouse (Reported)
Mouse BALB/c IgG2a, κ
Human SLP-76 SH2 domain Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645440
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560057 Rev. 2
Antibody Details
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H3

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.

560057 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
560057 Rev.2
Citations & References
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Development References (5)

  1. 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).
  2. 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
  3. Janssen E, Zhang W. Adaptor proteins in lymphocyte activation. Curr Opin Immunol. 2003; 15:269-276. (Biology). View Reference
  4. 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).
  5. Wu JN, Koretzky GA. The SLP-76 family of adapter proteins. Semin Immunol. 2004; 16:379-393. (Biology). View Reference
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560057 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.