Actin Protein - Lyophilized

Extracted from rabbit muscle to >99% and lyophilized for storage at room temperature.

Product: Ultra-Pure Actin

Catalog Number: 160101

Product Specifications:

Purity: >99%

Source: rabbit skeletal muscle

Molecular Weight: ~43 kDa

Form: desiccated powder (1 mg actin protein is supplied as 3 mg powder with extra mass attributed to trehalose, a lyoprotectant)

Buffer Conditions Upon Reconstitution: 2 mM Tris-HCl, 0.2 mM CaCl2, 0.2 mM ATP, 1 mM DTT, and 0.25 M Trehalose (pH 8.0)

Shipping: shipped at ambient temperatures

Storage Conditions: store in a cool, dry environment

Shelf Life: check product label for expiration date

Material

Ultra-Pure Actin is extracted from rabbit skeletal muscle using an optimized version of the method of Spudich and Watt (1971) and lyophilized by an adaptation of the method of Dráberová et al. (2010). The resulting actin protein is >99% pure (Figure 1) and >90% polymerization competent (Figure 2). Ultra-Pure Actin is supplied as a white powder. When reconstituted with ultra pure water to 3 mg/ml, the buffer conditions are 2 mM Tris-HCl, 0.2 mM CaCl2, 0.2 mM ATP, 1 mM DTT, and 0.25 M Trehalose, pH 8.0. Note that 1 mg actin protein is supplied as 3 mg powder (extra mass attributed to trehalose, a lyoprotectant), and reconstitution/dilution should be based on the actin protein concentration.

Activity and Applications

Ultra-Pure Actin will polymerize into filamentous F-actin when supplemented with KCl and MgCl2, and kept above its critical concentration. Ultra-Pure Actin is suitable for use in a variety of cell-free experimental applications, and polymerization activity is detectable in fluorescence microscopy assays, turbidity assays, and ATPase assays. Visit our protocols page for common actin polymerization protocols.

  • antibody generation
  • drug discovery by high-throughput screening
  • in vitro biochemical and biophysical approaches
  • structural analysis by X-ray crystallography and electron microscopy

Greater Purity at a Better Price

Ultra Pure Actin Purity

Figure 1:

Ultra-Pure Actin is >99% pure. Coomassie G250-stained protein gel of Ultra-Pure Actin separated by SDS-PAGE. The actin protein appears as a single species migrating at ~43 kDa. Molecular weight markers and loaded protein quantities are indicated.

Figure 2:

Ultra-Pure Actin is >90% polymerization-competent. Ultra-Pure Actin polymerized in the absence (-PB) or presence (+PB) of Actin Polymer Buffer (10X, Cat. No. 000103; 50 mM KCl and 2 mM MgCl2) followed by centrifugation at 48,000 rpm for 1 hour.  Pellet (P) and supernatant (S) fractions were collected and subjected to SDS-PAGE and Coomassie G250-staining. >90% of Ultra-Pure Actin was incorporated into filaments as determined by measuring the residual protein concentration in the supernatant fraction.

Ultra Pure Actin Polymerization