Non-Polymerizing Actin (alpha skeletal muscle actin)

Protein

Actin (α-Actin)

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Protein description

Non-Polymerizing Actin is a-skeletal muscle actin (42kD, 375 aa), enzymatically cleaved between Gly42/Val43. As a result of this cleavage an essential monomer-monomer interaction site is inactivated, rendering the actin unpolymerizable in the presence of Ca++ and e.g. 100mM KCl, conditions where normal actin polymerizes. 

Thus, NP-actin is a valuable tool for G-actin-ligand studies in the presence of Ca++ and KCl. On the other hand NP-actin remains polymerizable in the presence of Mg++, with a critical concentration of 3µM (barbed end). 

Actin compatible buffers preserve the structural integrity of non-polymerizing actin, while its cleavage products (approx. 36kD and 6kD) are only obtained under denaturing conditions, e.g. SDS-PAGE.
 

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Quantity per unit

2x 250 µg

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 rabbit skeletal muscle, GPC.

Protein concentration

Determined by the Biuret method.

Storage instructions

Avoid refreezing.     NP-Actin stored at –70°C upon arrival will be stable in performance for at least 6 months from the date of purchase. Solubilized G-actin is kept on ice and should be used within 1 week. 

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.

CAS no.

 51005-14-2