G-Actin/F-actin In Vivo Assay Biochem Kit (BK037)

Product Uses Include

• To study the effects of pharmaceutical compounds on the ratio of G-actin to F-actin.
• To study the effects of mutated cell lines versus their parent cell line for the change in ratio of G-actin to F-actin.
• To study the effects of physical alterations of environment on the ratio of G-actin to F-actin. 

Introduction
The most reproducible and accurate method of determining the amount of filamentous actin (F-actin) content versus free globular-actin (G-actin) content in a cell population is to use Western blot quantitation of F-actin and G-actin cellular fractions (1-4).  The general approach is to homogenize cells in F-actin stabilization buffer, followed by centrifugation to separate the F-actin from G-actin pool. The fractions are then separated by SDS-PAGE and actin is quantitated by Western blot. The final result gives the most accurate method of determining the ratio of F-actin incorporated into the cytoskeleton versus the G-actin found in the cytosol. This kit contains all the reagents needed to perform this assay.

Kit contents
The kit contains sufficient materials for 30-100 assays depending assay setup and includes reagents for positive and negative controls. The following components are included:

1. Lysis and F-actin stabilization buffer
2. ATP (Cat. # BSA04)
3. Protease inhibitor cocktail (Cat. # PIC02)
4. F-actin enhancing control solution
5. F-actin depolymerization control solution
6. Control G-actin Standard (Cat. # AKL99)
7. Actin polyclonal antibody (Cat. # AAN01)
8. SDS sample buffer (5 x)
9. DMSO
10. Manual with detailed protocols and extensive troubleshooting guide