Datasheet
Actin Working Buffer
| Catalog Number | Volume |
|---|---|
| 000102 | 5 X 1 ml |
Store at 4°C and protect from light
For research use only. Not for medical or diagnostic use.
Material
Actin Working Buffer is comprised of 5 mM Tris-HCl and 0.2 mM CaCl2 (pH 8.0).
Storage and Handling
Actin Working Buffer is stable at 4°C for 1 year.
Activity and Applications
Actin Working Buffer supports the stability and activity of purified actin in vitro when supplemented with 0.2 mM ATP and 1 mM DTT.
Polymerization Protocol
Dilute Ultra-Pure Actin (Cat. No. 160101) or Pure Actin (Cat. No. 190101) to 0.4 mg/ml with Actin Working Buffer supplemented with 0.2 mM ATP and 1 mM DTT. Incubate on ice for 1 hour followed by centrifugation at 14k rpm for 15 minutes at 4°C. Collect the supernatant and add 1/10 volume of Actin Polymer Buffer (10X; Cat. No. 000103; 500 mM KCl and 20 mM MgCl2) and ATP to 1 mM. Incubate at room temperature for 1 hour to polymerize. The resulting actin filaments can be stored at 4°C for 2 weeks with the addition of 0.05% sodium azide.
