PLCβ3 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). PLCβ3 is directly activated by Gα subunits related to Gαq, Gβγ heterodimers, and the small GTPase, Rac2. The regulation of PLCβ3 by Gq-coupled receptors is critical for normal cardiomyocyte function.1,2
Protein is > 89% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). PLCβ3 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 10-1234 of human PLCβ3.
- 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: Homo sapiens.
Sequence: PLCβ3 (residues 10-1234); lacks 10 residues from its N-terminus. The N-terminus retains a Ser-Gln-Ala tripeptide remaining from the affinity tag.
Calculated molecular weight: 138,367 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 80,220 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.