Description
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 protein tyrosine kinases and become enzymatically activated via phosphorylation. It exists as two isoforms: PLC-γ1, which is ubiquitously expressed, and PLC-γ2, which is found primarily in the hematopoietic system. PLC-γ2-null mice exhibit impaired B lymphocyte maturation and defects in Fc receptor functions. Phosphorylation of PLC-γ2 at tyrosines 753 and 759 (Y759) is required for activation of PLC-γ2 enzyme activity. PLC-γ2 phosphorylation at Y759 can be induced by stimulation of the B cell receptor in Ramos cells, the collagen receptor in platelets, and the T cell receptor in Jurkat cells, and it occurs downstream of Btk and BLNK in the signaling cascade of activated B cells. The K86-689.37.73 antibody recognizes PLC-γ2 phosphorylated at Y759 in the SH2-SH3 linker region.
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.
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. 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.
3. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. 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.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
