Actin-Trialkit Crystallography (alpha-actin, rabbit skeletal muscle)

Product name

Actin

Target species

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Triallkit description

Crystallization of actin binding proteins with G-actin is complicated by the tendency of actin to polymerize into filaments. An advantage for co-crystallization of ligand-actin complexes, or investigation of conformational changes upon complex formation in either molecule, is the use of a non-polymerizing actin. Non-Polymerizing Actin is alpha-skeletal muscle actin 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++, but not in the presence of Mg++. Thus without chemical modification of actin a non-polymerizing actin can be used for crystallography and also for biochemical analysis. The ability of NP-Actin to bind to other proteins remains essentially unimpaired.

The Actin-Trialkit Crystallography contains 1x1.0mg Non-Polymerizing Actin for co-crystallization trials or biochemical applications, a Trialkit Manual for safe & error-free handling of NP-Actin, and buffer for monomeric actin. Please refer to the Actin-Toolkit Crystallography for larger quantities of NP-Actin. We guarantee stable performance of the kit components when stored at -70°C upon arrival for 6 months.

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

1x1.0mg NP-Actin (lyophilized powder);
1x100ml G-Actin Buffer with ATP (all components supplied as powder); 
1xToolkit Handbook Crystallography;

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

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.

CAS no.

51005-14-2 (Actin)