Actin-Trialkit G-Actin Binding
alpha Actin, Cardiac Muscle (Bovine)
Cat. # T8200-01
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Applications
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Proof of ligands biological activity by G-actin binding;
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Functional analysis of ligands binding to G-actin;
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Analysis of pharmacological compounds, proteins or lipids altering ligand-G-actin-interaction;
Description
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Product name
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Actin
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Target species
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Cardiac muscle alpha actin, bovine
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Molecular mass
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42kDa
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Triallkit description
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Binding of ligands to G-actin proves the biological activity of the ligand and identifies the ligand to be a cardiac G-actin binding protein. The assays of this kit are based on the spin-down analysis of ligands bound to G-actin. For this purpose G-actin has been conjugated to Sepharose™ beads (ActinBeads) which are ready-to-use.
ActinBeads are pelleted at low centrifugal forces and ligands bound to G-actin will be specifically co-precipitated. SDS-PAGE analysis will reveal binding of G-actin binding ligands. The Trialkit is further used for assays ranging from the determination of binding stoichiometries using gelsolin as reference, up to variation of buffer conditions to possibly optimize G-actin binding.
The kit contains 1x250µl ActinBeads (50% slurry) prepared from rabbit skeletal muscle alpha actin (Cat. # T8020-01) or bovine cardiac alpha actin (Cat. # T8200-01) and buffers to keep actin functional. The Trialkit Manual describes the use of ActinBeads. We guarantee a stable performance of the kit components when stored at -70°C upon arrival for 6 months.
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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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1x250 µl ActinBeads (50% slurry, lyophilized Sepharose™ conjugated actin);
2x50 ml MonoMix (lyophilized G-actin ATP-buffer);
1x Trialkit Manual G-Actin Binding;
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Actin 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 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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CAS no.
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51005-14-2 (Actin)
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Sepharose is a registered trademark of GE Healthcare.
Further Information
Material and Safety Data Sheet
Protein Sequence on NCBI