PLCβ2 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). PLCβ2 is expressed in cells of hematopoietic origin. It is especially critical for chemotaxis of professional phagocytes such as neutrophils and platelet aggregation. PLCβ2 is directly activated by Gα subunits related to Gαq, Gβγ heterodimers, and the small GTPase, Rac2.1,2
Protein is > 89% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). PLCβ2 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 1-1181 of human PLCβ2.
- 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: Residues 1-1181 of PLCβ2. The N-terminus retains a Gly-Ala-Met-Asp-Pro-Glu-Phe peptide remaining from the affinity tag.
Calculated molecular weight: 134,427 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 98,210 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.