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Technical Data Sheet

PP1491

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PLCγ1 (Tyr-775), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
150 kDa

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Western blot analysis of PLCγ1 immunoprecipitates from human jurkat cells untreated (lanes 1 & 3) or treated with pervanadate (1 mM) for 30 min (lanes 2,4,5,6). Immunoprecipitation was performed with anti-PLCγ1 (PM1561). The blots were probed with anti-PLCγ1 (lanes 1 & 2) and anti-PLCγ1 (Tyr-775) (lanes 3-6). The latter antibody was used in the presence of phospho- PLCγ1 (Tyr-775) peptide (lane 5), or unrelated phosphotyrosine peptide (lane 6).

Application
Dilution


ELISA
1:2000


ICC
1:100


IP
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Fan, K. et al. (2012) J Gen Virol. 93(5):987. WB: Jukat cells
Ayuso-Sacido, A. et al. (2010) J Neuroonc. 97:323. WB: mouse neural stem cells
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Background
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). At least four families of PLCs have been identified: PLCβ, PLCγ, PLCδ, and PLCε. Phosphorylation is one of the key mechanisms that regulates the activity of PLC. PLCδ is activated by both receptor and nonreceptor tyrosine kinases. PLCγ1 forms a complex with EGF and PDGF receptors, which leads to phosphorylation at tyrosine 771, 783, and 1245. In addition, antigen receptor-induced activation of PLCγ1 leads to phosphorylation at both Tyr-775 and Tyr-783. These two sites are equally important for activation of enzymatic activity.


Background References
Chen, P. et al. (1996) J Cell. Biol. 134:689.
Obermeier, A. et al. (1996) EMBO J. 15:73.
Serrano, C.J. et al. (2005) J. Immunol. 174:6233.
Immunogen
PLCγ1 (Tyr-775) synthetic peptide (coupled to carrier protein) corresponding to amino acids around tyrosine 775 in human PLCγ1. This sequence has high homology to the conserved site in rat and mouse PLCγ1, and has low homology to PLCγ2.
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody was cross-adsorbed to a non-specific phospho-tyrosine peptide then affinity-purified using phospho-PLCγ1 (Tyr-775) peptide. The antibody detects a 150 kDa* protein in human Jurkat and A431 cells treated with pervanadate, but is not observed in untreated cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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