• Study the role of M44/47 oxidation on actin function
• Compare the in vitro functionality of oxidized vs non-oxidized actin in your experimental system
• Follow M44/47 oxidized actin polymerization in vitro by fluorescent enhancement

Regulation of actin oxidation at Met44/Met47 has been shown to destabilize F-actin in vivo and to play a key role in a growing number of cellular processes, including cytokinesis, axonal guidance, dendritic organization, synaptic development, heart and muscle development, and cell viability.

Pyrene-labeled rabbit muscle actin Cat. # AP05 has been n enzymatically oxidized at methionine 44 and 47 (-actin nomenclature) with the MICAL flavoprotein monooxygenase protein Cat. # {{SKU:MIC01|MIC01).

• Molecular weight ~43 kDa
• Formulation: supplied as a lyophilized powder
• Composition: when resuspended to 10 mg/ml, the buffer composition is: 5 mM Tris-HCl, pH 8.0, 0.2 mM CaCl2, 0.2 mM ATP, 5% (w/v) sucrose, and 1% (w/v) dextran

Protein purity is assessed by scanning densitometry of Coomassie Blue-stained protein on a 4-20% polyacrylamide gel. Purity is determined to be ≥95% pure.

The biological activity of pyrene-labeled oxidized actin has been determined by its ability to polymerize into filaments in vitro efficiently. At an actin concentration of 0.1 mg/ml, non-oxidized (reduced) pyrene actin exhibits a 7–12-fold increase in fluorescence (Ex: 360 nm, Em: 405 nm) upon polymerization, equivalent to >80% polymerization. Under the same conditions, oxidized actin is 40-55% as efficient, which is in agreement with published data.