Product Pathways - Ca / cAMP / Lipid Signaling
Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb #8713
|8713S||100 µl (10 western blots)||---||In Stock||---|
|8713||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Monkey||Endogenous||150||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Species predicted to react based on 100% sequence homology: Rat.
Specificity / Sensitivity
Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb recognizes endogenous levels of PLCγ1 protein only when phosphorylated at Ser1248.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser1248 of human PLCγ1 protein.
Western blot analysis of extracts from serum-starved A-431 and A549 cells, untreated (-) or treated (+) with hEGF #8916 (100 ng/mL, 15 min) or serum-starved NIH/3T3 cells, untreated (-) or treated (+) with hPDGF-BB #8912 (50 ng/mL, 15 min), using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb (upper) or PLCγ1 (D9H10) XP® Rabbit mAb #5690 (lower).
Immunoprecipitation (IP)/Western blot analysis of extracts from serum-starved HeLa cells, untreated (-) or treated (+) with TPA #4174 (100 nM, 15 min) prior to lysis in SDS (lanes 1 and 2) or IP lysis buffer (lane 3, TPA-treated only). IP Lysates were then subjected to immunoprecipitation with Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb (lane 4), PLCγ1 (D9H10) XP® Rabbit mAb #5690 (lane 5), or Normal Rabbit IgG #2729 (lane 6). The western blot was probed using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb. Lane 3 represents 10% input.
Immunohistochemical analysis of paraffin-embedded human colon (normal adjacent to tumor) using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb.
Immunohistochemical analysis of SignalSlide® Phospho-EGF Receptor IHC Controls #8102 [paraffin-embedded KYSE450 cell pellets untreated (left) or EGF-treated (right)] using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb.
Confocal immunofluorescent analysis of A-431 cells, serum starved (left) or treated with hEGF #8916 (100 ng/mL for 15 min) using Phospho-PLCγ1 (Ser1248) (D25A9) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Phosphoinositide-specific phospholipase C (PLC) plays a significant role in transmembrane signaling. In response to extracellular stimuli such as hormones, growth factors, and neurotransmitters, PLC hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to generate two secondary messengers: inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG) (1). At least four families of PLCs have been identified: PLCβ, PLCγ, PLCδ, and PLCε. Phosphorylation is one of the key mechanisms that regulate the activity of PLC. PLCγ is activated by both receptor and non-receptor tyrosine kinases (2). PLCγ forms a complex with EGF and PDGF receptors, which leads to the phosphorylation of PLCγ at Tyr771, 783, and 1245 (3). Phosphorylation by Syk at Tyr783 activates the enzymatic activity of PLCγ1 (4). PLCγ2 is engaged in antigen-dependent signaling in B cells and collagen-dependent signaling in platelets. Phosphorylation by Btk or Lck at Tyr753, 759, 1197, and 1217 is correlated with PLCγ2 activity (5,6).
Two mammalian PLCγ isoforms (γ1 and γ2) have been cloned and characterized (7,8). Like other PLC-family members, PLCγ1 and PLCγ2 contain calcium-binding (EF-hand, C2) and lipid-interacting (PH, EF-hand) domains necessary for their enzymatic activity and substrate recognition. Uniquely, PLCγ isoforms have additional, conserved SH2 and SH3 domains critical for their functions as signaling molecules and scaffolding proteins. Upon growth factor stimulation, PLCγ1 is recruited (via SH2 domains) to phosphotyrosine residues within the cytoplasmic tail of many RTKs where it serves as a substrate for the RTK and provides docking sites for additional proteins involved in RTK signaling (4-6,9-12). PLCγ1 and γ2 can also be activated downstream of receptors lacking intrinsic tyrosine kinase activity. This has been reported downstream of multiple G protein-coupled receptors and the T cell receptor in which tyrosine kinases of the Src, Syk, and Tec families serve to bind, phosphorylate, and activate PLCγ (reviewed in 13-15). Phosphorylation at tyrosine residues by both receptor and non-receptor tyrosine kinases results in robust activation of PLCγ1 activity, leading to generation of second messengers. In response to agonists, PLCγ1 is phosphorylated on Tyr783, Tyr711, and Tyr1253 (Tyr753, Tyr759, and Tyr1217 in PLCγ2) resulting in robust PI-4,5-P2 hydrolysis (4-6,9-12). Interestingly recent evidence suggests a role for tyrosine kinase-independent regulation of PLCγ in some systems. For example, in response to EGF, proline-rich regions of Akt interact with the SH3 domain of PLCγ1 resulting in association of the two enzymes, phosphorylation of PLCγ1 at Ser1248, and enhanced cellular motility (16). This finding demonstrates that PLCγ1 can function as a "scaffold" between RTKs and Akt, thereby establishing a mechanism by which the Akt signaling pathway cross-talks with tyrosine kinases. However, the mechanism and functional significance of phosphorylation at Ser1248 remains to be fully clarified, as it has also been shown that PKA-mediated phosphorylation at this site is inhibitory to PLCγ1 tyrosine phosphorylation and phospholipase activity in CD3-treated Jurkat cells (17), suggesting that Ser1248 may be an allosteric regulator of PLCγ1 activity.
- Singer, W.D. et al. (1997) Annu Rev Biochem 66, 475-509.
- Margolis, B. et al. (1989) Cell 57, 1101-7.
- Kim, H.K. et al. (1991) Cell 65, 435-41.
- Wang, Z. et al. (1998) Mol Cell Biol 18, 590-7.
- Watanabe, D. et al. (2001) J Biol Chem 276, 38595-601.
- Ozdener, F. et al. (2002) Mol Pharmacol 62, 672-9.
- Burgess, W.H. et al. (1990) Mol Cell Biol 10, 4770-7.
- Ohta, S. et al. (1988) FEBS Lett 242, 31-5.
- Rodriguez, R. et al. (2001) J Biol Chem 276, 47982-92.
- Humphries, L.A. et al. (2004) J Biol Chem 279, 37651-61.
- Kim, Y.J. et al. (2004) Mol Cell Biol 24, 9986-99.
- Sekiya, F. et al. (2004) J Biol Chem 279, 32181-90.
- Carpenter, G. and Ji, Q. (1999) Exp Cell Res 253, 15-24.
- Rebecchi, M.J. and Pentyala, S.N. (2000) Physiol Rev 80, 1291-335.
- Rhee, S.G. (2001) Annu Rev Biochem 70, 281-312.
- Wang, Y. et al. (2006) Mol Biol Cell 17, 2267-77.
- Park, D.J. et al. (1992) J Biol Chem 267, 1496-501.
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