Product Uses Include
This is a 10 x solution that contains 500 mM KCl, 20 mM MgCl2 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.
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Question 1: Once resuspended, can left-over actin polymerization buffer be stored at 4°C to maintain activity?
Answer 1: Upon reconstitution (2.0 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’s experiments (This buffer is stable for 30 min on ice) and aliquot the remainder into 100 μl 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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