Product Uses Include
Bovine cardiac muscle actin is supplied as a lyophilized powder. When one vial is reconstituted in 1 ml of Actin Monomer Buffer: 5.0 mM Tris-HCI pH 8.0, 0.2 mM CaCl2, 0.2 mM ATP and 0.5 mM DTT, it is at a final concentration of 1.0 mg/ml.
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are > 99% pure.
The biological activity is determined in three ways. The first method utilizes pyrene muscle actin as a fluorescent indicator of polymerization (use a 1:10 ratio of AP07 to AD99) . The second involves sedimentation of actin polymer by high speed centrifugation. The third method measures the integrity of the monomer by the specific inhibition of DNase activity. The biological activity is > 90%.
|Lu, Yi Ju et al.||Arabidopsis calcium-dependent protein kinase 3 regulates actin cytoskeleton organization and immunity||Nature Communications||2020||ISSN 2041-1723|
|Seervai, Riyad N.H. et al.||The Huntingtin-interacting protein SETD2/HYPB is an actin lysine methyltransferase||Science Advances||2020||ISSN 2375-2548|
|Sandberg, Alexander et al.||Fibrillation and molecular characteristics are coherent with clinical and pathological features of 4-repeat tauopathy caused by MAPT variant G273R||Neurobiology of Disease||2020||ISSN 1095-953X|
|Mahmud, Zabed et al.||Structure and proteolytic susceptibility of the inhibitory C-terminal tail of cardiac troponin I||Biochimica et Biophysica Acta (BBA) - General Subjects||2019|
|Yang, Kai Chien et al.||Deficiency of nuclear receptor interaction protein leads to cardiomyopathy by disrupting sarcomere structure and mitochondrial respiration||Journal of molecular and cellular cardiology||2019||ISSN 1095--8584|
|Da’as, Sahar I. et al.||Hypertrophic cardiomyopathy-linked variants of cardiac myosin-binding protein C3 display altered molecular properties and actin interaction||Biochemical Journal||2018||ISSN 1470-8728|
|Manickam, Manoj et al.||Exploration of flexible phenylpropylurea scaffold as novel cardiac myosin activators for the treatment of systolic heart failure||European Journal of Medicinal Chemistry||2017||ISSN 1768-3254|
|Cuellar, Jorge et al.||Assisted protein folding at low temperature: Evolutionary adaptation of the antarctic fish chaperonin CCT and its client proteins||Biology Open||2014||ISSN 2046-6390|
|Butler, Suzanne C. et al.||Inhibitory effects of pectenotoxins from marine algae on the polymerization of various actin isoforms||Toxicology in Vitro||2012||ISSN 0887-2333|
|Orlova, Albina et al.||The N-Terminal Domains of Myosin Binding Protein C Can Bind Polymorphically to F-Actin||Journal of Molecular Biology||2011|
|Passarelli, Chiara et al.||Susceptibility of isolated myofibrils to in vitro glutathionylation: Potential relevance to muscle functions||Cytoskeleton||2010||ISSN 1949--3592|
|Debold, Edward P. et al.||Human actin mutations associated with hypertrophic and dilated cardiomyopathies demonstrate distinct thin filament regulatory properties in vitro||Journal of Molecular and Cellular Cardiology||2010||ISSN 0022--2828|
Question 1: What is the isotype composition of the cardiac actin from bovine heart muscle?
Answer 1: Bovine cardiac actin (Cat. # AD99) is composed of 84% a-cardiac and 16% a-skeletal actin isoforms.
Question 2: How do I measure actin polymerization with the pyrene assay when there is no available pyrene conjugate for cardiac actin?
Answer 2: To examine the polymerization of unlabeled cardiac actin (Cat. # AD99), please click here for a polymerization protocol that uses an excess of the unlabeled [cardiac] actin plus 10% of pyrene-labeled skeletal muscle actin. The pyrene muscle actin will not polymerize on its own at the concentration used in this assay, so the reaction is dependent on unlabeled cardiac actin (Cat. # AD99) for F-actin formation. In this way, the pyrene-labeled muscle actin is taken up and polymerized to serve as a reporter for polymerization of the unlabeled actin that is present at a much greater concentration.
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