Alpha-actinin protein: rabbit skeletal muscle

Alpha-actinin protein: rabbit skeletal muscle
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Product Uses Include

  • Discover and characterize α-actinin binding proteins
  • Positive control of actin binding assays
  • Positive control for actin bundling assays

Material
α-actinin has been purified from rabbit skeletal muscle. The protein is supplied as a lyophilized powder and upon reconstitution, will be in the following buffer: 4 mM Tris-HCl pH 7.6, 4 mM NaCl, 20 μM EDTA, 1% (w/v) sucrose, and 0.2% (w/v) dextran. The lyophilized product is stable at -70°C for at least 2 years. AT01 should be resuspended in nanopure water containing 1 mM β-mercaptoethanol or DTT to the desired working concentration. 

Purity
Purity is determined by scanning densitometry of the protein on an SDS-PAGE gel. α-actinin is >90% pure.


at01gel

Figure 1: α-actinin protein purity determination. 20 µg of AT01 was run on an SDS-PAGE gel and stained with coomassie blue.

Biological Activity
At 2.5 µM (0.25 mg/ml) α-actinin and 
25 µM (1.0 mg/ml) F-actin at pH 7.0 and 24°C, approximately 70% of the α-actinin will co-sediment with F-actin after centrifugation at 150,000 x g for 1 h.

For product Datasheets and MSDSs please click on the PDF links below.   For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

AuthorTitleJournalYearArticle Link
Bashirzadeh, Yashar et al.Encapsulated actomyosin patterns drive cell-like membrane shape changesiScience2022
Hsiao, Bo Yuan et al.Human costars family protein abracl modulates actin dynamics and cell migration and associates with tumorigenic growthInternational Journal of Molecular Sciences2021ISSN 1422-0067
Bashirzadeh, Yashar et al.Encapsulated Actomyosin Patterns Drive Cell-Like Membrane Shape ChangesSSRN Electronic Journal2021Article Link
Park, Jinho et al.Crowding tunes the organization and mechanics of actin bundles formed by crosslinking proteinsFEBS Letters2021ISSN 1873--3468
Rodríguez-Pérez, Fernando et al.Ubiquitin-dependent remodeling of the actin cytoskeleton drives cell fusionDevelopmental Cell2021ISSN 1878-1551
Liao, Kuo An et al.Characterizing the actin-binding ability of Zasp52 and its contribution to myofibril assemblyPLoS ONE2020ISSN 1932-6203
Gilmore, Jamie L. et al.AFM Investigation of the Organization of Actin Bundles Formed by Actin-Binding ProteinsJournal of Surface Engineered Materials and Advanced Technology2013ISSN 2161--4881
Van Der Gucht, Jasper et al.Stress release drives symmetry breaking for actin-based movementProceedings of the National Academy of Sciences of the United States of America2005ISSN 0027-8424
Harris, Elizabeth S. et al.The Mouse Formin, FRLα, Slows Actin Filament Barbed End Elongation, Competes with Capping Protein, Accelerates Polymerization from Monomers, and Severs FilamentsJournal of Biological Chemistry2004ISSN 0021-9258
Lu, Shajia et al.New N-RAP-binding partners alpha-actinin, filamin and Krp1 detected by yeast two-hybrid screening: implications for myofibril assemblyJournal of cell science2003ISSN 0021--9533
Maul, Raymond S. et al.EPLIN regulates actin dynamics by cross-linking and stabilizing filamentsJournal of Cell Biology2003ISSN 0021--9525
Salmikangas, Paula et al.Myotilin, the limb-girdle muscular dystrophy 1A (LGMD1A) protein, cross-links actin filaments and controls sarcomere assemblyHuman Molecular Genetics2003ISSN 0964-6906
Karakesisoglou, Iakowos et al.An Epidermal Plakin That Integrates Actin and Microtubule Networks at Cellular JunctionsJournal of Cell Biology2000ISSN 0021--9525
Waterman-Storer, Clare et al.Microtubules remodel actomyosin networks in Xenopus egg extracts via two mechanisms of F-actin transportJournal of Cell Biology2000ISSN 0021-9525

 

Question 1: What is the source of the alpha-actinin Cytoskeleton sells?

Answer 1:  Cytoskeleton’s alpha-actinin (Cat. # AT01) is purified from rabbit skeletal muscle and has a purity of >85%.

 

Question 2: Can this alpha-actinin be used for actin bundling experiments?

Answer 2: Yes, Cytoskeleton’s alpha-actinin (Cat. # AT01) is well-suited for actin bundling experiments using skeletal muscle actin, it will not work with non-muscle isoforms of actin.  Briefly, α-actinin is tested for biological activity by a co-sedimentation assay with Factin.  At 2.5 μM skeletal muscle α-actinin and 25 μM actin in the form of F-actin, >70% α-actinin will co-sediment with F-actin. The reaction is carried out at pH 7.0 for 30 minutes at 24°C (at pH 8.0 α-actinin will not bind).  Spin the mixture at 14,000 x g for 10 minutes.  Probe a western blot with antibodies against alpha-actinin and actin.  An SDS-PAGE gel is loaded with supernatant and pellet samples from alpha-actinin + actin and actin alone. After running the gel to separate samples it is stained with coomassie blue and then analyzed for band intensity at 43 and 116 kDal.  

 

 

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