Question 1: What is the source of the alpha-actinin Cytoskeleton sells?
Answer 1: Cytoskeleton&rsquo, s alpha-actinin (Cat. # AT01) is purified from rabbit skeletal muscle and has a purity of >, 85%.
Question 2: Can this alpha-actinin be used for actin bundling experiments?
Answer 2: Yes, Cytoskeleton&rsquo, s alpha-actinin (Cat. # AT01) is well-suited for actin bundling experiments using skeletal muscle actin, it will not work with non-muscle isoforms of actin. Briefly, &alpha, -actinin is tested for biological activity by a co-sedimentation assay with Factin. At 2.5 &mu, M skeletal muscle &alpha, -actinin and 25 &mu, M actin in the form of F-actin, >, 70% &alpha, -actinin will co-sediment with F-actin. The reaction is carried out at pH 7.0 for 30 minutes at 24°, C (at pH 8.0 &alpha, -actinin will not bind). Spin the mixture at 14, 000 x g for 10 minutes. Probe a western blot with antibodies against alpha-actinin and actin. An SDS-PAGE gel is loaded with supernatant and pellet samples from alpha-actinin + actin and actin alone. After running the gel to separate samples it is stained with coomassie blue and then analyzed for band intensity at 43 and 116 kDal.
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