Actin protein (bovine cardiac muscle)

Product Uses Include

Identification and characterization of cardiac actin binding proteins
In vitro cardiac actin polymerization studies

Material:
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:
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are > 99% pure.

Biological Activity:
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%.

Click Here for Detailed Methods

About

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

For a guide to performing actin polymerizations with this actin product please click here.

Citations

Author	Title	Journal	Year	Article Link
Martin, Jose L. et al.	Actin Isoform Composition and Binding Factors Fine-Tune Regulatory Impact of Mical Enzymes	International Journal of Molecular Sciences	2023	ISSN 1422--0067
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	ISSN 0304--4165
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	ISSN 0022--2836
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

Faqs

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.

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