PLCγ1 is a calcium-dependent lipase that hydrolyzes phosphatidylinositol 4,5- bisphosphate (PIP2) to produce the second messengers, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). PLCγ1 is activated upon phosphorylation by a wide variety of receptor tyrosine kinases and soluble tyrosine kinases to control proliferation and differentiation.1,2
Protein is > 94% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). PLCγ1 was heterologously expressed in High 5 insect cells as a histidine-tagged protein and purified by a combination of affinity, ion exchange, and gel exclusion chromatographies. The tag was removed prior to final purification leaving a Ser-Gln-Ala tripeptide at the N-terminus followed by residues 1-1219 of rat PLCγ1. Purified PLCγ1 lacks 71 residues from its C-terminus that are not conserved. This truncation improves purity and solubility without affecting lipase activity.
- Kadamur G, et al., Mammalian phospholipase C. Annu Rev Physiol 75, 15.1-15.28 (2013).
- Gresset A, et al., Mechanism of phosphorylation-induced activation of phospholipase C-γ isozymes. J Biol Chem 285, 35836-47 (2010).
Species: Rattus norvegicus – 94% identity to Homo sapiens.
Sequence: PLCγ1 (residues 1-1219); lacks 71 residues from its C-terminus that are not conserved. The N-terminus retains a Ser-Gln-Ala tripeptide remaining from the affinity tag.
Calculated molecular weight: 140,469 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 179,010 M-1cm-1 (assuming all cysteines remain reduced).
Amount: 20 μg.
Concentration: 0.2 mg/ml (determined by absorbance at 280 nm).
Buffer: 20 mM Hepes pH 7.5, 150 mM NaCl, 5% glycerol and 1 mM DTT.
Form: Frozen liquid.
Storage and stability
Storage: -80 °C. After initial thaw, aliquot and store at -80 °C until further use.
Stability: Stable for up to 12 months at -80 °C and up to 2 days at 4 °C. Repeated freeze-thaw cycles not recommended.
Notes: Please see accompanying sheet describing activity.