Actin protein ( >99% pure): rabbit skeletal muscle

Actin protein (>99% pure): rabbit skeletal muscle
$0.00

Product Uses Include

  • Identification and characterization of muscle actin binding proteins
  • In vitro actin polymerization studies
  • Antibody standard for Western blot analysis

 

Material
Actin protein has been purified from rabbit skeletal muscle. AKL99 actin is greater than 99% pure and AKL95 is greater than 95% pure. Muscle actin has an approximate molecular weight of 43 kDa. Rabbit muscle actin is supplied as a white lyophilized powder. The lyophilized protein when stored desiccated to <10% humidity at 4°C is stable for 6 months. When reconstituted in distilled water to 10 mg/ml, the protein is in the following buffer: 5 mM Tris-HCl pH 8.0, 0.2 mM CaCl2, 0.2 mM ATP, 5% sucrose, and 1% dextran.

Purity
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% polyacrylamide gel. AKL99 consists of > 99% pure muscle actin while AKL95 is > 95% pure (see Figure 1).

aklgel

Figure 1:  Purities of rabbit skeletal muscle actin protein. 100 µg of > 99% pure (AKL99) and > 95% pure (AKL95) rabbit skeletal muscle actin were run on SDS-PAGE gels and stained with coomassie blue. The arrow indicates actin protein, the arrowhead an α-actinin contaminant (115 kDa). The minor impurities in the purified actins are predominantly actin binding proteins such as α-actinin and gelsolin.

Biological Activity
The biological activity of muscle actinis determined by its ability to efficiently polymerize into filaments (F-actin) in vitro and separate from unpolymerized components in a spin down assay. Stringent quality control ensures that AKL99 produces > 90% F-actin and AKL95 produces > 80% F-actin in this assay.

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
Haciosmanoglu Aldogan, Ebru et al.Depolymerization of actin filaments by Cucurbitacin I through binding G-actinFood Science & Nutrition2024ISSN 2048--7177
Paladini, Serena et al.Measurement and Characterization of the Electrical Properties of Actin FilamentsInternational Journal of Molecular Sciences 2024, Vol. 25, Page 54852024ISSN 1422--0067
McCormick, Laura E. et al.Multi-monoubiquitylation controls VASP-mediated actin dynamicsJournal of Cell Science2024ISSN 1477-9137
Chand, Abhishu et al.CdSe/ZnS Quantum Dots’ Impact on In Vitro Actin DynamicsInternational Journal of Molecular Sciences 2024, Vol. 25, Page 41792024ISSN 1422--0067
Morales, E. Angelo et al.Mitotic spindle positioning protein (MISP) preferentially binds to aged F-actinJournal of Biological Chemistry2024ISSN 0021--9258
Smith, Amanda J. et al.Design and Construction of a Multi-Tiered Minimal Actin Cortex for Structural Support in Lipid Bilayer ApplicationsACS Applied Bio Materials2024ISSN 2576-6422
Chen, Xiaobing et al.Crowding alters F-actin secondary structure and hydrationCommunications Biology 2023 6:12023ISSN 2399--3642
Rivera Alvarez, José et al.The kinesin Kif21b regulates radial migration of cortical projection neurons through a non-canonical function on actin cytoskeletonCell reports2023ISSN 2211--1247
Wang, Xue et al.Loss of ORP3 induces aneuploidy and promotes bladder cancer cell invasion through deregulated microtubule and actin dynamicsCellular and Molecular Life Sciences2023ISSN 1420-9071
Domínguez-García, Pablo et al.Micro-mechanical response and power-law exponents from the longitudinal fluctuations of F-actin solutionsSoft Matter2023ISSN 1744-6848
Martin, Annabell et al.A general strategy to develop fluorogenic polymethine dyes for bioimagingNature Chemistry 2023 16:12023ISSN 1755--4349
Nakamura, Hideki et al.ActuAtor, a Listeria-inspired molecular tool for physical manipulation of intracellular organizations through de novo actin polymerizationCell reports2023ISSN 2211--1247
Henry, Conor M. et al.SYK ubiquitination by CBL E3 ligases restrains cross-presentation of dead cell-associated antigens by type 1 dendritic cellsCell reports2023ISSN 2211--1247
Ashour, Dina J. et al.Zasp52 strengthens whole embryo tissue integrity through supracellular actomyosin networksDevelopment2023ISSN 0950--1991
Ji, Xinmiao et al.Intermittent F-actin Perturbations by Magnetic Fields Inhibit Breast Cancer MetastasisResearch2023ISSN 2639-5274
Domínguez-García, Pablo et al.Micro-mechanical response and power-law exponents from the longitudinal fluctuations of F-actin solutionsSoft Matter2023ISSN 1744-6848
Henty-Ridilla, Jessica L.Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence (TIRF) MicroscopyJoVE (Journal of Visualized Experiments)2022ISSN 1940--087X
Sasanpour, Mehrzad et al.Reconstituting and Characterizing Actin-Microtubule Composites with Tunable Motor-Driven Dynamics and MechanicsJoVE (Journal of Visualized Experiments)2022ISSN 1940--087X
Cheng, Xiaohang et al.Reconstitution of Phase-Separated Signaling Clusters and Actin Polymerization on Supported Lipid BilayersFrontiers in Cell and Developmental Biology2022ISSN 2296-634X
Bashirzadeh, Yashar et al.Encapsulated actomyosin patterns drive cell-like membrane shape changesiScience2022
Morales, E. Angelo et al.Mitotic Spindle Positioning (MISP) is an actin bundler that selectively stabilizes the rootlets of epithelial microvilliCell Reports2022
Sheung, Janet Y. et al.Motor-driven advection competes with crowding to drive spatiotemporally heterogeneous transport in cytoskeleton compositesFrontiers in Physics2022
Wu, Yueh-Fu O. et al.The CLIP-170 N-terminal domain binds directly to both F-actin and microtubules in a mutually exclusive mannerJournal of Biological Chemistry2022ISSN 0021-9258
Arbore, C. et al.α-catenin switches between a slip and an asymmetric catch bond with F-actin to cooperatively regulate cell junction fluidityNature Communications2022ISSN 2041-1723
Würtz, Martin et al.Modular assembly of the principal microtubule nucleator γ-TuRCNature Communications2022ISSN 2041-1723
Rimoli, Caio Vaz et al.4polar-STORM polarized super-resolution imaging of actin filament organization in cellsNature Communications2022ISSN 2041-1723
Lin, Lin et al.Phase separation-mediated condensation of Whirlin-Myo15-Eps8 stereocilia tip complexCell Reports2021ISSN 2211-1247
Giampazolias, Evangelos et al.Secreted gelsolin inhibits DNGR-1-dependent cross-presentation and cancer immunityCell2021ISSN 1097-4172
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
Lee, Gloria et al.Myosin-driven actin-microtubule networks exhibit self-organized contractile dynamicsScience Advances2021ISSN 2375-2548
Peris-Moreno, Dulce et al.Ube2l3, a partner of murf1/trim63, is involved in the degradation of myofibrillar actin and myosinCells2021ISSN 2073-4409
Colombo, Jessica et al.A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energeticsNature Communications2021ISSN 2041-1723
Iv, Francois et al.Insights into animal septins using recombinant human septin octamers with distinct SEPT9 isoformsJournal of cell science2021ISSN 1477-9137
Yang, Haibin et al.Complex structures of rsu1 and pinch1 reveal a regulatory mechanism of the ilk/ pinch/parvin complex for f-actin dynamicseLife2021ISSN 2050-084X
Rodríguez-Pérez, Fernando et al.Ubiquitin-dependent remodeling of the actin cytoskeleton drives cell fusionDevelopmental Cell2021ISSN 1878-1551
Ricketts, Shea N. et al.Triggering Cation-Induced Contraction of Cytoskeleton Networks via MicrofluidicsFrontiers in Physics2020ISSN 2296-424X
Lin, Shan Shan et al.Dynamin-2 Regulates Postsynaptic Cytoskeleton Organization and Neuromuscular Junction DevelopmentCell Reports2020ISSN 2211-1247
Cheng, Yu Shu et al.The load dependence and the force-velocity relation in intact myosin filaments from skeletal and smooth musclesAmerican Journal of Physiology - Cell Physiology2020ISSN 1522-1563
Kumar, Ashok et al.Actin R256 Mono-methylation Is a Conserved Post-translational Modification Involved in TranscriptionCell Reports2020ISSN 2211-1247
Liao, Kuo An et al.Characterizing the actin-binding ability of Zasp52 and its contribution to myofibril assemblyPLoS ONE2020ISSN 1932-6203
Hsiao, An Shan et al.An intrinsically disordered protein interacts with the cytoskeleton for adaptive root growth under stressPlant Physiology2020ISSN 1532-2548
Wu, Hongpo et al.Alloxan disintegrates the plant cytoskeleton and suppresses Mlo-mediated powdery mildew resistancePlant and Cell Physiology2020ISSN 1471-9053
Sonal et al.Myosin-II activity generates a dynamic steady state with continuous actin turnover in a minimal actin cortexJournal of Cell Science2019ISSN 1477-9137
Francis, Madison L. et al.Non-monotonic dependence of stiffness on actin crosslinking in cytoskeleton compositesSoft Matter2019ISSN 1744-6848
Stavusis, Janis et al.Novel mutations in MYBPC1 are associated with myogenic tremor and mild myopathyAnnals of Neurology2019ISSN 1531-8249
Shashi, Vandana et al.Heterozygous variants in MYBPC1 are associated with an expanded neuromuscular phenotype beyond arthrogryposisHuman Mutation2019ISSN 1098-1004
Gardini, Lucia et al.A protocol for single molecule imaging and tracking of processive myosin motorsMethodsX2019ISSN 2215-0161
Piroli, Gerardo G. et al.Identification of novel protein targets of DMF modification in neurons and astrocytes reveals actions independent of Nrf2 stabilizationMolecular and Cellular Proteomics2019ISSN 1535-9484
Anderson, Sylas J. et al.Filament Rigidity Vies with Mesh Size in Determining Anomalous Diffusion in CytoskeletonBiomacromolecules2019ISSN 1526-4602
Li, Zhiwei et al.Homer Tetramer Promotes Actin Bundling Activity of DrebrinStructure2019ISSN 1878-4186
Liang, Xiaoling et al.Transgelin 2 is required for embryo implantation by promoting actin polymerizationFASEB Journal2019ISSN 1530-6860
Balta, Emre et al.Spatial oxidation of L-plastin downmodulates actin-based functions of tumor cellsNature Communications2019ISSN 2041-1723
Laitila, Jenni et al.A nebulin super-repeat panel reveals stronger actin binding toward the ends of the super-repeat regionMuscle and Nerve2019ISSN 1097-4598
Steinz, Maarten M. et al.Oxidative hotspots on actin promote skeletal muscle weakness in rheumatoid arthritisJCI Insight2019ISSN 2379-3708
Antoku, Susumu et al.ERK1/2 Phosphorylation of FHOD Connects Signaling and Nuclear Positioning Alternations in Cardiac LaminopathyDevelopmental Cell2019ISSN 1878-1551
Kommaddi, Reddy Peera et al.Aβ mediates F-actin disassembly in dendritic spines leading to cognitive deficits in alzheimer’s diseaseJournal of Neuroscience2018ISSN 1529-2401
Zeng, Menglong et al.Reconstituted Postsynaptic Density as a Molecular Platform for Understanding Synapse Formation and PlasticityCell2018ISSN 1097-4172
Papadaki, Maria et al.Diabetes with heart failure increases methylglyoxal modifications in the sarcomere, which inhibit functionJCI insight2018ISSN 2379-3708
Ricketts, Shea N. et al.Co-Entangled Actin-Microtubule Composites Exhibit Tunable Stiffness and Power-Law Stress RelaxationBiophysical Journal2018ISSN 1542-0086
Ahmed, Zubair M. et al.Inframe deletion of human ESPN is associated with deafness, vestibulopathy and vision impairmentJournal of Medical Genetics2018ISSN 1468-6244
Weise-Cross, Laura et al.Actin polymerization contributes to enhanced pulmonary vasoconstrictor reactivity after chronic hypoxiaAmerican Journal of Physiology - Heart and Circulatory Physiology2018ISSN 1522-1539
Khatra, Harleen et al.Hedgehog Antagonist Pyrimidine–Indole Hybrid Molecule Inhibits Ciliogenesis through Microtubule DestabilisationChemBioChem2018ISSN 1439-7633
Heier, Jonathon A. et al.Measuring protein binding to F-actin by co-sedimentationJournal of Visualized Experiments2017ISSN 1940-087X
Ordija, Christine M. et al.Free actin impairs macrophage bacterial defenses via scavenger receptor MARCO interaction with reversal by plasma gelsolinAmerican Journal of Physiology - Lung Cellular and Molecular Physiology2017ISSN 1522-1504
Kubiniok, Peter et al.Time-resolved phosphoproteome analysis of paradoxical RAF activation reveals novel targets of ERKMolecular and Cellular Proteomics2017ISSN 1535-9484
Garagounis, Constantine et al.Microcompartmentation of cytosolic aldolase by interaction with the actin cytoskeleton in ArabidopsisJournal of Experimental Botany2017ISSN 1460-2431
Goryunov, D et al.Microtubule-actin cross-linking factor 1: domains, interaction partners, and tissue-specific functionsMethods in Enzymology2016Article Link
Belyy, Alexander et al.Actin activates Pseudomonas aeruginosa ExoY nucleotidyl cyclase toxin and ExoY-like effector domains from MARTX toxinsNature Communications2016ISSN 2041-1723
Yamanaka, Daisuke et al.Phosphatidylinositol 3-kinase-associated protein (PI3KAP)/XB130 crosslinks actin filaments through its actin binding and multimerization properties In Vitro and enhances endocytosis in HEK293 cellsFrontiers in Endocrinology2016ISSN 1664-2392
Zheng, Lili et al.Correction to Characterization and regulation of an additional actin-filamentbinding site in large isoforms of the stereocilia actin-bundling protein espin [Journal of Cell Science (2015), 127, 1306-1317]Journal of Cell Science2015ISSN 1477-9137
Walrant, Astrid et al.Triggering actin polymerization in xenopus egg extracts from phosphoinositide-containing lipid bilayersMethods in Cell Biology2015ISSN 0091-679X
Falzone, Tobias T. et al.Entangled F-actin displays a unique crossover to microscale nonlinearity dominated by entanglement segment dynamicsSoft Matter2015ISSN 1744-6848
Chan, Eddie et al.The acetylenic tricyclic bis(cyano enone), TBE-31 inhibits non-small cell lung cancer cell migration through direct binding with actinCancer Prevention Research2014ISSN 1940-6215
Luo, Shen et al.Taurine chloramine-induced inactivation of cofilin protein through methionine oxidationFree Radical Biology and Medicine2014ISSN 1873-4596
Maiden, Stephanie L. et al.Specific conserved C-terminal amino acids of caenorhabditis elegans HMP-1/α-catenin modulate f-actin binding independently of vinculinJournal of Biological Chemistry2013ISSN 0021-9258
Butler, Suzanne C. et al.Inhibitory effects of pectenotoxins from marine algae on the polymerization of various actin isoformsToxicology in Vitro2012ISSN 0887-2333
Jensen, Mikkel Herholdt et al.Effects of basic calponin on the flexural mechanics and stability of F-actinCytoskeleton2012ISSN 1949--3592
Windhorst, Sabine et al.Functional role of inositol-1,4,5-trisphosphate-3-kinase-A for motility of malignant transformed cellsInternational Journal of Cancer2011ISSN 1097--0215
Zucchini, Daniela et al.Kinetic and spectroscopic characterization of the putative monooxygenase domain of human MICAL-1Archives of Biochemistry and Biophysics2011ISSN 0003-9861
Zhang, Tong et al.Growth-Arrest-Specific protein 2 inhibits cell division in xenopus embryosPLoS ONE2011ISSN 1932-6203
Choi, Chang Hoon et al.Expression of Actin-interacting Protein 1 Suppresses Impaired Chemotaxis of Dictyostelium Cells Lacking the Na+-H+ Exchanger NHE1Molecular Biology of the Cell2010ISSN 1059-1524
Romagnoli, Silvia et al.Cytosolic proteins from tobacco pollen tubes that crosslink microtubules and actin filaments in vitro are metabolic enzymesCytoskeleton2010ISSN 1949-3584
Eiseler, Tim et al.Protein kinase D controls actin polymerization and cell motility through phosphorylation of cortactinJournal of Biological Chemistry2010ISSN 0021-9258
Del Duca, Stefano et al.Effects of post-translational modifications catalysed by pollen transglutaminase on the functional properties of microtubulesand actin filamentsBiochemical Journal2009ISSN 1470-8728
Sawyer, Gregory M. et al.Disease-associated Substitutions in the Filamin B Actin Binding Domain Confer Enhanced Actin Binding Affinity in the Absence of Major Structural Disturbance: Insights from the Crystal Structures of Filamin B Actin Binding DomainsJournal of Molecular Biology2009ISSN 0022-2836
Norman, Holly et al.Myofibrillar protein and gene expression in acute quadriplegic myopathyJournal of the Neurological Sciences2009ISSN 0022-510X
Clark, Alice R. et al.Skeletal dysplasias due to filamin A mutations result from a gain-of-function mechanism distinct from allelic neurological disordersHuman Molecular Genetics2009ISSN 0964-6906
Chellaiah, Meenakshi A. et al.Phosphorylation of a Wiscott-Aldrich syndrome protein-associated signal complex is critical in osteoclast bone resorptionJournal of Biological Chemistry2007ISSN 0021-9258
Allingham, John S. et al.A Structural Basis for Regulation of Actin Polymerization by PectenotoxinsJournal of Molecular Biology2007ISSN 0022-2836
Arora, Pamela D. et al.Gelsolin Mediates Collagen Phagocytosis through a Rac-dependent StepMolecular Biology of the Cell2004ISSN 1059-1524
Ishikawa, Takashi et al.Subdomain organization of the Acanthamoeba myosin IC tail from cryo-electron microscopyProceedings of the National Academy of Sciences of the United States of America2004ISSN 0027-8424
Goryunov, Dmitry et al.Studying cytolinker proteinsMethods in Cell Biology2004ISSN 0091-679X
Upadhyaya, Arpita et al.Probing polymerization forces by using actin-propelled lipid vesiclesProceedings of the National Academy of Sciences of the United States of America2003ISSN 0027-8424
Loomis, Patricia A. et al.Espin cross-links cause the elongation of microvillus-type parallel actin bundles in vivo2003PMID 14657236
Balcer, Heath I. et al.Coordinated Regulation of Actin Filament Turnover by a High-Molecular-Weight Srv2/CAP Complex, Cofilin, Profilin, and Aip12003PMID 14680631
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
Humphries, Christine L. et al.Direct regulation of Arp2/3 complex activity and function by the actin binding protein coronin2002ISSN 0021--9525
Sagot, Isabelle et al.An actin nucleation mechanism mediated by Bni1 and ProfilinNature Cell Biology 2002 4:82002ISSN 1476--4679
Engqvist-Goldstein, Åsa E.Y. et al.The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitroJournal of Cell Biology2001ISSN 0021-9525
McGhie, E. J. et al.Cooperation between actin-binding proteins of invasive Salmonella: SipA potentiates SipC nucleation and bundling of actinThe EMBO Journal2001ISSN 1460--2075

Question 1:  What is the best way to store actin proteins to insure maximum stability and shelf-life?

Answer 1:  Cytoskeleton provides all of our actin proteins as lyophilized powders so that they can be shipped at room temperature.  Upon receipt, the lyophilized powders should be stored at 4°C in a sealed container with desiccant.  It is important to monitor the freshness of the desiccant and insure that it continues to absorb moisture to protect the lyophilized actins.  With proper storage, the lyophilized actins are guaranteed to be stable for 6 months from the date of purchase.  Alternatively, actins can be immediately resuspended at the concentration recommended, aliquoted, snap-frozen in liquid nitrogen and stored at -70°C.  The frozen aliquots will be stable for 6 months.  When thawing frozen aliquots, it is important to thaw in a room temperature water bath.

 

Question 2:  What is the best way to store F-actin after polymerizing?

Answer 2:  G-actin is stable for two days at 4°C and requires a divalent cation, pH 6.5 - 8.0 and ATP for stability.  F-actin is stable and can be stored at 4°C for 1-2 weeks.  F-actin requires ATP (0.2 mM) and Mg2+ (2 mM) for stability and is unstable below pH 6.5 and above pH 8.5.  F-actin is not stable to freezing.  F-actin can be transferred to a variety of buffers (e.g. HEPES, phosphate, etc) without detrimental effects.  We recommend the addition of antibacterial agents such as 100 μg/ml ampicillin and 10 μg/ml chloramphenicol when storing F-actin at 4°C.

 

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