|H M||Endogenous||Rabbit IgG|
Flow cytometric analysis of human whole blood using BLNK (D3P2H) XP® Rabbit mAb (PE Conjugate) and co-stained with CD19-FITC (left) and CD3-APC (right). CD19+ B cells are positive for BLNK, whereas CD3+ T cells are negative. Analysis was performed on cells in the lymphocyte gate.Learn more about how we get our images.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Account for isotype-matched controls for monoclonal antibodies or species matched IgG for polyclonal antibodies.
Reference: Chow S, Hedley D, Grom P, Magari R, Jacobberger JW, Shankey TV (2005) Whole blood fixation and permeabilization protocol with red blood cell lysis for flow cytometry of intracellular phosphorylated epitopes in leukocyte subpopulations. Cytometry A 67(1), 4–17.
posted November 2008
revised September 2013
Protocol Id: 384
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
BLNK (D8P2H) XP® Rabbit mAb (PE Conjugate) recognizes endogenous levels of total BLNK protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg282 of human BLNK protein.
This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated BLNK (D8P2H) XP® Rabbit mAb #36438.
B cell linker protein (BLNK), also known as SLP-65 or BASH, is an adaptor molecule that plays key roles in B cell activation and B cell antigen receptor (BCR) engagement. BLNK acts at the interface between BCR-associated Syk and downstream signaling cascades (1,2). BLNK has multiple SH2 binding motifs (YXXP) at its amino terminus and an SH2 domain at its carboxy terminus. After BCR ligation, BLNK is phosphorylated by Syk at multiple YXXP motifs including Tyr72, Tyr84, Tyr96, and Tyr178 (1). These phosphorylated motifs provide docking sites for signaling molecules, such as BTK, PLCγ, and Vav. These signaling molecules bind to BLNK through their SH2 domains and together activate downstream signaling pathways (3,4). Through its SH2 domain, BLNK can also interact with tyrosine-phosphorylated targets, such as HPK1, thereby recruiting them to the BCR complex for signaling (5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc.
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|48947S||100 µl (50 tests)||N/A|
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