Actin-Toolkit Crystallography

Protein

Non-Polymerizing Actin

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Toolkit description

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++, while it is fully polymerizable in the presence of Mg++. Also in the presence of e.g. 100mM KCl, NP-Actin does not polymerize unless 2mM MgCl2 are added to the solution.

Crystallization of actin binding proteins with G-actin is complicated by the polymerization of actin. An advantage for crystallization of actin and co-crystallization of ligand-actin complexes is the use of Non-Polymerizing Actin.  The Toolkit is furthermore used for biophysical methods investigating conformational changes upon complex formation in actin or the actin ligand.

Without chemical modification of actin, Non-Polymerizing Actin can be used for crystallography, biophysical or biochemical analysis. The ability of NP-Actin to bind to other proteins remains essentially unimpaired.

For initial tests, we offer the Actin-Trialkit Crystallography (Cat. # T8040-01). The Actin-Toolkit Crystallography contains 6x 1 mg Non-Polymerizing Actin prepared from purified rabbit skeletal muscle actin. A handbook with a detailed description for a safe, error-free handling of NP-Actin is provided with the kit.

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

6x 1.0 mg NP-Actin (lyophilized powder);
1x 1000 ml G-Actin Buffer with ATP (all components supplied as powder); 
2x 1.0 ml Mg-Exchange buffer
1x Toolkit Handbook Crystallography;

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 100µl ultrapure water to obtain a 1mg/ml solution.

DOL

-

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. 

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