Actin-Toolkit SPR (alpha skeletal muscle actin)

Protein

Actin (α-skeletal muscle Actin)

Origin

Skeletal muscle alpha actin, rabbit

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. Furthermore, the specific actin binding strongly indicates that the ligand is correctly folded and structurally intact.

The Actin-Toolkit SPR provides a unique method to precisely analyze ligand-F-actin interactions by Surface Plasmon Resonance techniques on Streptavidin chips. To overcome the general problem in SPR - which is that the actin filaments clog the capillaries of the device - short actinfilaments with Ankerprotein^TM on the fast polymerizing barbed ends are used for for coating. The actin filaments protrude from the surface of the chip and permit the introduction of ligands. After the actin filaments become attached to the surface of the chip, ligands / analytes can be introduced to investigate binding to F-actin. 

The Actin-Toolkit SPR was developed to analyze 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 it is suitable to investigate effects of pharmacological compounds on ligand-actin complexes, as well as further proteins, lipids etc. to ligand-actin complexes. 

The kit contains rabbit skeletal muscle actin (Cat. # 8010-03), Ankerprotein^TM, FAB-EP, GAP-EP (SPR actin buffers for F-and G-actin). A handbook introduces and guides through the experimental work with actin in SPR.

Please note, that Streptavidin coated chips are required and not included. The setup for the compounds to be investigated has to be determined individually.

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

4x250 µg Actin (lyophilized powder);
4x10 µg Ankerprotein^TM (lyophilized powder);
1x PolyMix (actin polymerization buffer, 10x);
2x250 ml FAB-P;
1x250 ml GAB-P;
1x1.0 ml PolyMix;
1x100mM Phalloidin (500µl);
1xToolkit Handbook SPR;

Buffers

FAB-P: 10mM Imidazol-Cl pH 7.4, 0.4mM ATP, 2mM MgCl2 and 0.0005% v/v Surfactant P20.
GAB-P: 2mM Tris-Cl pH 8.2, 0.4mM ATP, 0.5mM DTT, 0.1mM CaCl2 and 0.0005% v/v Surfactant P20.

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 skeletal muscle, rabbit; 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.