Actin-Toolkit F-Actin Binding
α-actin from bovine cardiac muscle
Cat. #: 8100-01
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Applications
- Proof of the biological activity of a ligand by actin binding;
- Cosedimentation analysis of ligands binding to F-actin;
- Testing of actin filament bundling activity or network formation by ligands;
- Analysis of pharmacological compounds, proteins or lipids altering ligand-actin-interaction;
Description
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Protein
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Actin (α-cardiac Actin)
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Origin
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Cardiac muscle alpha actin, bovine
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Molecular mass
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42kDa
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Toolkit description
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Binding of ligands to filamentous actin is used as a functional proof to state the biological activity of a ligand and characterizes the ligand to be an actin binding protein. Specific actin binding indicates that the ligand is correctly folded and structurally intact. The Actin-Toolkit F-Actin Binding is a well-established and commonly used method to characterize ligand-actin interaction: at high speed centrifugation ligands binding to individual actin filaments are co-sedimented.
At low speed spinning a selective co-sedimentation of actin filament bundles or networks is achieved. Low speed assays instantly characterize ligands to induce actin suprastructures.
The Actin-Toolkit F-Actin Binding was developed to characterize the actin-binding properties of ligands (native or recombinant proteins, recombinant fragments etc.) and to support the detection even of cryptic actin-binding sites. Further suitable applications are in vitro compound analysis of the ligand-actin interactions by investigating effects of pharmacological compounds on ligand-actin complexes, as well as the introduction of further ligands (proteins, lipids etc.) to ligand-actin complexes.
The kit contains 4x1 mg bovine cardiac muscle actin (Cat. # 8100-02), buffers to keep actin functional and to prevent denaturing. A handbook introduces and guides through the experimental work with actin in F-actin Binding Assays.
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Purity
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>99% by scanning densitometry.
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Properties
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Form
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Lyophilized, ready-to-use.
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Kit content
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4x1 mg Actin (lyophilized powder);
4x50 ml MonoMix (lyophilized G-actin ATP-buffer);
1x1 ml PolyMix, 10x stock (lyophilized F-actin ATP-buffer, pH 7.4);
1x1 ml PolyMix 6, 10x stock (lyophilized F-actin ATP-buffer, pH 6.0);
1x1 ml PolyMix 8, 10x stock (lyophilized F-actin ATP-buffer, pH 8.0);
1x0.5 ml MgCl2 (solution)
1xToolkit Handbook F-Actin Binding;
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Buffer
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2mM Tris-Cl pH 8.2, 0.4mM ATP, 0.5mM DTT, 0.1mM CaCl2, 1mM NaN3 and 0.3% disaccharides, when reconstituted with 1.0 ml ultrapure water to obtain a 1mg/ml solution.
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Related products
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For dilution of G-actin or exchange of ligand buffer into actin compatible buffer, MonoMix (Cat. #: 5100-0*) may be used. In the absence of nucleators, the polymerization of G-actin is initiated by PolyMix (Cat. #: 5000-0*).
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Purification notes
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Purified from bovine cardiac muscle, GPC.
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Protein concentration
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Determined by the Biuret method.
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Storage instructions
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We guarantee stable performance of the kit components for 6 months when stored at -70°C upon arrival. Solubilized G-actin is kept on ice and should be used within 1 week. Avoid refreezing.
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Shipping conditions
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At ambient temperature. Upon delivery store at -70°C.
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Remarks
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For Use in Research only. Not for Use in Human or Veterinary Diagnostical or Therapeutical Applications.
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Further Information
Material and Safety Data Sheet
Protein Sequence on NCBI