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

Actin protein (>99% pure): rabbit skeletal muscle
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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
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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
Bashirzadeh, Yashar et al.Encapsulated Actomyosin Patterns Drive Cell-Like Membrane Shape ChangesSSRN Electronic Journal2021Article Link
Lee, Gloria et al.Myosin-driven actin-microtubule networks exhibit self-organized contractile dynamicsScience Advances2021ISSN 2375-2548
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
Gübeli, Raphael J. et al.In Vitro-Evolved Peptides Bind Monomeric Actin and Mimic Actin-Binding Protein Thymosin-β4ACS Chemical Biology2021ISSN 1554-8937
Iv, Francois et al.Insights into animal septins using recombinant human septin octamers with distinct SEPT9 isoformsJournal of cell science2021ISSN 1477-9137
Colombo, Jessica et al.A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energeticsNature Communications2021ISSN 2041-1723
Anderson, S. J. et al.Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networksSoft Matter2021ISSN 1744-6848
Kučera, Ondřej et al.Anillin propels myosin-independent constriction of actin ringsNature Communications2021ISSN 2041-1723
Lee, Gloria et al.Active cytoskeletal composites display emergent tunable contractility and restructuringSoft Matter2021ISSN 1744-6848
Hsiao, An Shan et al.An intrinsically disordered protein interacts with the cytoskeleton for adaptive root growth under stressPlant Physiology2020ISSN 1532-2548
Kumar, Ashok et al.Actin R256 Mono-methylation Is a Conserved Post-translational Modification Involved in TranscriptionCell Reports2020ISSN 2211-1247
Wu, Hongpo et al.Alloxan disintegrates the plant cytoskeleton and suppresses Mlo-mediated powdery mildew resistancePlant and Cell Physiology2020ISSN 1471-9053
Jarsch, Iris K. et al.A direct role for SNX9 in the biogenesis of filopodiaJournal of Cell Biology2020ISSN 1540-8140
Ricketts, Shea N. et al.Triggering Cation-Induced Contraction of Cytoskeleton Networks via MicrofluidicsFrontiers in Physics2020ISSN 2296-424X
Rai, Amrita et al.The mechanism of activation of the actin binding protein EHBP1 by Rab8 family membersNature Communications2020ISSN 2041-1723
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
Lin, Shan Shan et al.Dynamin-2 Regulates Postsynaptic Cytoskeleton Organization and Neuromuscular Junction DevelopmentCell Reports2020ISSN 2211-1247
Liao, Kuo An et al.Characterizing the actin-binding ability of Zasp52 and its contribution to myofibril assemblyPLoS ONE2020ISSN 1932-6203
Liang, Xiaoling et al.Transgelin 2 is required for embryo implantation by promoting actin polymerizationFASEB Journal2019ISSN 1530-6860
Li, Zhiwei et al.Homer Tetramer Promotes Actin Bundling Activity of DrebrinStructure2019ISSN 1878-4186
Anderson, Sylas J. et al.Filament Rigidity Vies with Mesh Size in Determining Anomalous Diffusion in CytoskeletonBiomacromolecules2019ISSN 1526-4602
Steinz, Maarten M. et al.Oxidative hotspots on actin promote skeletal muscle weakness in rheumatoid arthritisJCI Insight2019ISSN 2379-3708
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
Balta, Emre et al.Spatial oxidation of L-plastin downmodulates actin-based functions of tumor cellsNature Communications2019ISSN 2041-1723
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
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
Sieck, Gary C. et al.Mechanisms underlying TNFα-induced enhancement of force generation in airway smooth musclePhysiological Reports2019ISSN 2051-817X
Stavusis, Janis et al.Novel mutations in MYBPC1 are associated with myogenic tremor and mild myopathyAnnals of Neurology2019ISSN 1531-8249
Lin, Shan-Shan et al.F-actin Bundle Sedimentation AssayBio-Protocol2019ISSN 2331--8325
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
Cheng, Yu Shu et al.Cleavage of loops 1 and 2 in skeletal muscle heavy meromyosin (HMM) leads to a decreased functionArchives of Biochemistry and Biophysics2019ISSN 1096-0384
Antoku, Susumu et al.ERK1/2 Phosphorylation of FHOD Connects Signaling and Nuclear Positioning Alternations in Cardiac LaminopathyDevelopmental Cell2019ISSN 1878-1551
Kumar, Dhivya et al.Microvillar and ciliary defects in zebrafish lacking an actin-binding bioactive peptide amidating enzymeScientific Reports2018ISSN 2045-2322
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
Ahmed, Zubair M. et al.Inframe deletion of human ESPN is associated with deafness, vestibulopathy and vision impairmentJournal of Medical Genetics2018ISSN 1468-6244
Ricketts, Shea N. et al.Co-Entangled Actin-Microtubule Composites Exhibit Tunable Stiffness and Power-Law Stress RelaxationBiophysical Journal2018ISSN 1542-0086
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
Khatra, Harleen et al.Hedgehog Antagonist Pyrimidine–Indole Hybrid Molecule Inhibits Ciliogenesis through Microtubule DestabilisationChemBioChem2018ISSN 1439-7633
Gardini, Lucia et al.Dissecting myosin-5B mechanosensitivity and calcium regulation at the single molecule levelNature Communications2018ISSN 2041-1723
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
Yeyati, Patricia L. et al.KDM3A coordinates actin dynamics with intraflagellar transport to regulate cilia stabilityJournal of Cell Biology2017ISSN 1540-8140
Shepardson, Kelly M. et al.Induction of antiviral immune response through recognition of the repeating subunit pattern of viral capsids is toll-like receptor 2 dependentmBio2017ISSN 2150-7511
Garagounis, Constantine et al.Microcompartmentation of cytosolic aldolase by interaction with the actin cytoskeleton in ArabidopsisJournal of Experimental Botany2017ISSN 1460-2431
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
Heier, Jonathon A. et al.Measuring protein binding to F-actin by co-sedimentationJournal of Visualized Experiments2017ISSN 1940-087X
Kubiniok, Peter et al.Time-resolved phosphoproteome analysis of paradoxical RAF activation reveals novel targets of ERKMolecular and Cellular Proteomics2017ISSN 1535-9484
Goryunov, D et al.Microtubule-actin cross-linking factor 1: domains, interaction partners, and tissue-specific functionsMethods in Enzymology2016Article Link
Gurmessa, Bekele et al.Entanglement Density Tunes Microscale Nonlinear Response of Entangled ActinMacromolecules2016ISSN 1520-5835
Ambrosi, Cinzia et al.Connexin43 forms supramolecular complexes through non-overlapping binding sites for drebrin, tubulin, and ZO-1PLoS ONE2016ISSN 1932-6203
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
Belyy, Alexander et al.Actin activates Pseudomonas aeruginosa ExoY nucleotidyl cyclase toxin and ExoY-like effector domains from MARTX toxinsNature Communications2016ISSN 2041-1723
Walrant, Astrid et al.Triggering actin polymerization in xenopus egg extracts from phosphoinositide-containing lipid bilayersMethods in Cell Biology2015ISSN 0091-679X
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
Fukumoto, Miki et al.α-Actinin-4 enhances colorectal cancer cell invasion by suppressing focal adhesion maturationPLoS ONE2015ISSN 1932-6203
Falzone, Tobias T. et al.Entangled F-actin displays a unique crossover to microscale nonlinearity dominated by entanglement segment dynamicsSoft Matter2015ISSN 1744-6848
Luo, Shen et al.Taurine chloramine-induced inactivation of cofilin protein through methionine oxidationFree Radical Biology and Medicine2014ISSN 1873-4596
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
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
Chacón-Martínez, Carlos Andrés et al.The switch-associated protein 70 (SWAP-70) bundles actin filaments and contributes to the regulation of F-actin dynamicsJournal of Biological Chemistry2013ISSN 0021-9258
Lu, Wenshu et al.Nesprin interchain associations control nuclear sizeCellular and Molecular Life Sciences2012ISSN 1420-682X
Butler, Suzanne C. et al.Inhibitory effects of pectenotoxins from marine algae on the polymerization of various actin isoformsToxicology in Vitro2012ISSN 0887-2333
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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.