Actin-Toolkit F-Actin Binding (alpha cardiac actin)

Protein

Actin (α-cardiac Actin)

Origin

Cardiac muscle alpha actin, bovine

Molecular mass

42kDa

Toolkit description

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.

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

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;

Buffer

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.

Related products

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*). 

Purification notes

Purified from bovine cardiac muscle, GPC.

Protein concentration

Determined by the Biuret method.

Storage instructions

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.     

Shipping conditions 

At ambient temperature. Upon delivery store at -70°C.

Remarks

For Use in Research only. Not for Use in Human or Veterinary Diagnostical or Therapeutical Applications.