MICAL-Oxidized (Pyrene labeled) Actin Protein (>95% pure): Rabbit Skeletal Muscle

MICAL-Oxidized Pyrene Actin Protein (>95% pure): Rabbit Skeletal Muscle (250 micrograms)
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MICAL interacts with F-actin and uses NADPH as a cofactor to oxidize actin at Met44 and Met47 (b-actin nomenclature). Functionally, oxidation of Met44 has a profound effect on actin polymerization because the residue resides in the D-loop of subdomain 2 of the protein, which is critical for actin subunit contacts; thus, upon oxidation, Met44 becomes negatively charged and interferes with actin monomer-monomer interaction and promotes F-actin severing and depolymerization. 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.  As part of the MOXtrue™ product line, Pyrene labeled rabbit skeletal muscle actin protein (MICAL-oxidized) (MXAP95) has been enzymatically oxidized at methionines 44 and 47  with the MICAL flavoprotein monoxygenase protein. Purified MICAL-oxidized (pyrene labeled) actin has reduced susceptibility to subtilisin A cleavage at M47/G48 by > 90%, and has also been validated for downstream applications such as actin polymerization assays. 

To learn more about using MICAL-oxidized Actin as a research tool see our datasheet 

To learn more about the MICAL/MsrB/Actin physiological redox system see our Newsletter 

Each lot of purified protein is quality controlled  to provide high batch to batch consistency, see COA documents.


Actin Polymerization of Oxidized Pyrene Actin Versus Native Pyrene Actin

Pyrene-labeled actin (Cat # AP05) and MICAL-oxidized (pyrene labeled) actin (Cat. # MXAP95) were diluted to 0.1 mg/ml (2.3 μM) or 0.2 mg/ml (4.6 μM) (see method). Samples were then incubated with 2x polymerization buffer.  Upon actin polymerization fluorescence was detected with a spectrophotometer. A.U. = arbitrary units

Click here for a detailed method 

MXAP95_actin_polymerization

For more information contact:  signalseeker@cytoskeleton.com

Associated Products:

MOXtrue™ 6xHis MICAL-1 Protein (Cat. # MIC01)

MOXtrue™ 6xHis MsrB2 Protein (Cat. # MB201)

MOXtrue™ MICAL-oxidized Actin (Cat. # MXA95)

Rabbit Skeletal Muscle Actin (Cat. # AKL95)

Pyrene Labeled Rabbit Skeletal Muscle Actin (Cat.# AP05)

For product Datasheets and MSDSs please click on the PDF links below.

Certificate of Analysis:  Lot 001

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If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com