PLCβ1 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). PLCβ1 is mainly expressed in the cerebral cortex and hippocampus, where the enzyme regulates neuronal activity. PLCβ1 is directly activated by Gα subunits related to Gαq and Gβγ heterodimers.1,2
Protein is > 98% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). PLCβ1 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 Gly-Ala-Met-Asp-Pro-Glu-Phe peptide at the N-terminus followed by residues 1-1216 of rat PLCβ1.
- Kadamur G, et al., Mammalian phospholipase C. Annu Rev Physiol 75, 15.1-15.28 (2013).
- Lyon A.M, et al., Structural Insights into Phospholipase C-β Function. Mol Pharmacol 83, 488-500 (2013).
Species: Rattus norvegicus – 97% identity to Homo sapiens.
Sequence: Residues 1-1216 of PLCβ1. The N-terminus retains a Gly-Ala-Met-Asp-Pro-Glu-Phe peptide remaining from the affinity tag.
Calculated molecular weight: 139,092 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 92,600 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.