Detection of Human Phospho-PLC‑ gamma 2 (Y759) by Western Blot.
Western blot shows lysates of Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 100 µM Pervanadate (PV) for 10 minutes or 10 µg/mL Goat Anti-Human IgM μ Chain Antigen Affinity-purified Polyclonal Antibody (Catalog # G-105-C) for 2 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-PLC‑ gamma 2 (Y759) Monoclonal Antibody (Catalog # MAB7377) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-PLC‑ gamma 2 (Y759) at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-PLC‑ gamma 2 (Y759) in Ramos Human Cell Line.
PLC‑ gamma 2 phosphorylated at Y759 was detected in immersion fixed Ramos human Burkitt's lymphoma cell line treated (upper panel) or untreated (lower panel) with pervanadate for 5 minutes using Mouse Anti-Human Phospho-PLC‑ gamma 2 (Y759) Monoclonal Antibody (Catalog # MAB7377) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: PLC-gamma 2
The Phospholipase C family consists of 13 isozymes within six subfamilies, PLC-δ, -beta, -gamma, -epsilon, -zeta, and -eta. PLC-gamma 2 (Phospholipase C gamma-2) contains 2 SH2 and 1 SH3 domains and is primarily limited to cells of hematopoietic lineage. PLC-gamma 2 is activated by receptor tyrosine kinases in response to growth factors, neurotransmitters, and hormones, and downstream through Lck kinase-dependent phosphorylation at Y753 and Y759. Activated PLC-gamma 2 catalyzes the hydrolysis of phosphatidylinositol 4, 5-biphosphate to produce the second messengers inositol 1, 4, 5-triphosphate (IP3) and diacylglycerol (DAG). IP3 mobilizes the release of calcium while DAG activates protein kinase C. PLC-gamma 2 is involved in collagen induced signaling in platelets and antigen-dependent signaling in B-lymphocytes. Human PLC-gamma 2 shares 94% and 95% sequence identity overall with mouse and rat PLC-gamma 2, respectively, and 100% sequence identity within the peptide immunogen.
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