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Preparation And Storage
Recommended Assay Procedures
This PLC-γ2-specific antibody conjugate may be used with conjugates of anti-PLC-γ2 (pY759) mAb K86-689.37 to distinguish the expression of total versus phosphorylated PLC-γ2.
This antibody conjugate is suitable for intracellular staining of human whole blood and mouse splenocytes using the BD Phosflow™ Lyse/Fix Buffer and the BD Phosflow™ Perm Buffer II.
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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- 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® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The Phospholipase C (PLC) isozymes hydrolyze phosphatidyl inositol biphosphate to inositol triphosphate and diacylglycerol. The former causes release of calcium from the endoplasmic reticulum, while the latter is an activator of Protein Kinase C. Within the PLC family, PLC-γ is the only member that contains SH2 and SH3 domains. These domains enable it to interact with receptor tyrosine kinases and become enzymatically activated via phosphorylation. It exists as two isoforms: 1) PLC-γ1, which is ubiquitously expressed, and 2) PLCγ2, found primarily in the lymphoid system. PLC-γ is essential for growth factor-induced cell motility and mitogenesis. Overexpression of PLC-γ is evident in several forms of cancer, and it has been identified as a key mediator of PDGF-dependent cellular transformation. Thus regulation of PLC-γ activity by growth factors is involved in cell growth and transformation.
Although the immunogen for generation of the K86-1161 monoclonal antibody was a phosphorylated peptide, peptide blocking studies demonstrated that the mAb recognizes PLC-γ2 regardless of phosphorylation status. This antibody was raised to a unique region of PLC-γ2 and is predicted not to crossreact with PLC-γ1
Development References (5)
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Kim YJ, Sekiya F, Poulin B, Bae YS, Rhee SG. Mechanism of B-cell receptor-induced phosphorylation and activation of phospholipase C-γ2. Mol Cell Biol. 2004; 24(22):9986-9999. (Biology).
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Knoll M, Yanagisawa Y, Simmons S, et al. The non-Ig parts of the vpreB and λ5 proteins of the surrogate light chain play opposite roles in the surface representation of the precursor b cell receptor. J Immunol. 2012; 188(12):6010-6017. (Clone-specific: Flow cytometry). View Reference
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Ozdener F, Dangelmaier C, Ashby B, Kunapuli SP, Daniel JL. Activation of phospholipase Cγ2 by tyrosine phosphorylation.. Mol Pharmacol. 2002; 62(3):672-679. (Biology).
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Wang D, Feng J, Wen R, et al. Phospholipase Cγ2 is essential in the functions of B cell and several Fc receptors. Immunity. 2000; 13:25-35. (Biology).
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Wen R, Jou S-T, Chen Y, Hoffmeyer A, Wang D. Phospholipase Cγ2 is essential for specific functions of FcεR and FcγR . J Immunol. 2002; 169:6743-6752. (Biology).
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.