Shop By Categories
- Tubulin Protein (7)
- Actin Protein (2)
- Buffers (3)
- Gift Shop (4)
To get updates on discounts and sale items join our list.
Journal Club
Read the latest news in the microtubule industry.
Refresh | This website puresoluble.com/protocols/actin-pelleting-assay/ is currently offline. Cloudflare's Always Online™ shows a snapshot of this web page from the Internet Archive's Wayback Machine. To check for the live version, click Refresh. |
At relatively high centripetal forces, filamentous F-actin will pellet while globular G-actin will remain suspended in solution. This biophysical property can be leveraged in probing for actin-associated proteins and assessing actin polymerization competency. This protocol outlines how to fractionate actin protein solutions by centrifugation and probe the resulting pellet and supernatant fractions.
Follow Steps 1-6 of the Polymerizing Actin Protein protocol to generate F-actin filaments.
Spin the F-actin solution at 48,000 rpm (100,000 x g) for 1 hour at 26°C in an ultracentrifuge rotor (i.e. TLA-100).
Collect the supernatant fraction and add an equal volume of 2X Sample Buffer.
Gently wash the pellet twice with Actin Working Buffer.
Resuspend the pellet in 1X Sample Buffer to a final volume matching that of the supernatant fraction.
Analyze the pellet and supernatant fractions by SDS-PAGE. Polymerization reactions devoid of KCl and MgCl2 can serve as a negative control.
Read the latest news in the microtubule industry.
For general, purchasing, or technical questions, email [email protected].
PurSolutions or its third party tools uses cookies to improve your experience on this website. Some of the cookies we use are essential for parts of the website to operate. By Accepting, Closing this banner or browsing further, you agree to the use of cookies. For more information on our use of cookies, please review our