PLCδ1 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). In addition to the calcium co-factor used to hydrolyze PIP2, PLCδ1 also binds several calcium ions through its C2 domain. These additional calcium ions anchor PLCδ1 to negatively charged membranes and account for the exquisite sensitivity of this PLC to fluctuation in calcium.1,2
Protein is > 93% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). PLCδ1 was heterologously expressed in E. coli 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-Asn-Ala tripeptide at the N-terminus followed by residues 1-755 of human PLCδ1.
- Kadamur G, et al., Mammalian phospholipase C. Annu Rev Physiol 75, 15.1-15.28 (2013).
- Cocco L, et al., Phosphoinositide-specific phospholipase C in health and disease. J Lipid Res 56(10), 1853–1860 (2015).
Species: Homo sapiens.
Sequence: Residues 1-755 of PLCδ1. The N-terminus retains a Ser-Asn-Ala tripeptide remaining from the affinity tag.
Calculated molecular weight: 85,806 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 91,790 M-1cm-1 (assuming all cysteines remain reduced).
Amount: 100 μg.
Concentration: 1 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.