Identification of calcium binding proteins from brain
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A procedure was worked out for purification and identification of calcium-binding proteins from bovine brain using Ca2+-dependent, reversible binding to a hydrophobic support, phenyl-Sepharose, as the method of isolation. These proteins could be visualized during and after their separation by running them on non-denaturing polyacrylamide gels, blotting to Zeta-probe paper, and autoradiographing with45Ca2+. About 24 polypeptides could be seen in this fraction on SDS (Laemmli) gels and about 8–10 native, Ca2+-binding proteins could be seen on non-denaturing gels and on blots of their 45Ca2+ autoradiographs. Some of these proteins could be purified further by chromatography on DEAE-Sephacel and still retain their45Ca2+-binding activity.
Key WordsCalcium binding protein isolation purification
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