Actin-Toolkit G-Actin Binding (alpha skeletal muscle actin)

Protein

Actin (α-Actin)

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Toolkit description

The interaction of G-actin with lignds is highly specific. Thus, a positive proof of this interaction strongly indicates that G-actin binding is a biological function of the ligand. All assays of the Actin-Toolkit G-Actin Binding are performed with G-actin coupled to Sepharose™ (ActinBeads), with the option to use gelsolin as reference protein. 

The handbook included in the kit guides through different experimental approaches. In combination with the buffers provided the ActinBeads are kept functional and are prevented from denaturing during assay performance. The Actin Toolkit G-Actin Binding contains 4x250µl G-actin (50% slurry) conjugated to Sepharose™ to be spun at low centrifugal forces. Under these conditions ligands bound to G-actin will be co-precipitated with actin. SDS-PAGE will reveal binding of G-actin binding ligands.  

The Actin-Toolkit G-Actin Binding is further used for assays ranging from the determination of binding stoichiometries up to the variation of buffer conditions to possibly optimize G-actin binding. The kit is available with Sepharose™ conjugated actin from rabbit skeletal (Cat. # 8020-01) . Further components are gelsolin, and buffers to keep actin functional. A handbook introduces to G-actin and guides through the experimental use of ActinBeads. We guarantee stable performance of all kit components when stored at -70°C upon arrival for at least 6 months. 

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

4x250µl ActinBeads (lyophilized sepharose conjugated actin);
4x50ml MonoMix (lyophilized G-actin ATP-buffer);
1x0.5ml 100mM CaCl2 (steril filtered);
1x15µg Gelsolin (mammalian); 
5x Syringes & Needles; 
1x Toolkit Handbook G-Actin Binding;

Actin 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. 

Purification notes

Purified from rabbit skeletal 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.