Actin Polymerization Buffer (10x stock)

Product Uses Include

• Polymerization of G-actin into F-actin

Material
This is a 10 x solution that contains 500 mM KCl, 20 mM MgCl_2, 50 mM guanidine carbonate pH 7.5, and 10 mM ATP in 100 mM Tris, pH 7.5. It is used to add to G-actin solutions in A-buffer (General actin buffer (Cat. # BSA01), 0.2 mM ATP (Cat. # BSA04) and 0.5 mM DTT) to induce polymerization into F-actin.

Question 1: Once resuspended, can left-over actin polymerization buffer be stored at 4°, C to maintain activity?

Answer 1: Upon reconstitution (1.8 ml of 100 mM Tris-HCl pH 7.5), the actin polymerization buffer (Cat. # BSA02) will be at 10X strength.  Set aside the volume needed for that day&rsquo, s experiments (This buffer is stable for 30 min on ice) and aliquot the remainder into 100 &mu, l volumes, snap frozen in liquid nitrogen, and stored at -70°, C. These stocks are stable for 6 months at -70°, C.

Question 2: After resuspension, does the actin polymerization buffer need to be used in a certain amount of time before losing activity?

Answer 2: The actin polymerization buffer (Cat. # BSA02) will be stable for 30 min after thawing the previously frozen aliquots (or using freshly prepared). Keep on ice until ready to use.  We do not recommend re-freezing aliquots.

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