Products

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-Polymerizaing 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 ablility of NP-Actin

to bind to other proteins remains essentially unimpared.

For initial tests, we offer the Actin-Trialkit Crystallography (Cat. # T8040-01).

The Actin-Toolkit Crystallography contains 10x1 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

10x1.0mg NP-Actin (lyophilized powder)

1x1000ml G-Actin Buffer with ATP (all components supplied as powder)

2x1.0ml Mg-Exchange buffer 

1xToolkit 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 1.0 ml ultrapure water to obtain a 1mg/ml

solution.

DOL

>0.5

Purification notes

Purified from rabbit skeletal muscle, GPC.

DOL

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.