Alexa Fluor® 647 Mouse anti-SLP-76 (pY128)
Clone J141-668.36.58 (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing) Mouse, Rat (Predicted)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human SLP-76
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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 phosphorylated tyrosine 128 (Y128) brings the Rho family guanine nucleotide exchange factor Vav1 and the Nck adapter protein, which binds to p21-activated kinase (PAK1) and Wiskott-Aldrich syndrome protein (WASP), into the activation complex. Vav1, PAK1, and WASP may mediate TCR-stimulated actin cytoskeletal rearrangement.
The J141-688.36.58 monoclonal antibody recognizes the phosphorylated Y128 of activated SLP-76.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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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.
- 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).
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
This antibody conjugate is suitable for intracellular staining of human whole blood (using BD Phosflow™ Lyse/Fix Buffer, Cat. No. 558049) and peripheral blood mononuclear cells (using BD Cytofix™ Fixation Buffer, Cat. No. 554655). Any of the 3 BD Phosflow™ Perm Buffers may be used.