Figure 1).

Identification and purification of high molecular weight calmodulin (CaM)-binding protein (HMWCaMBP) from cardiac muscle. Heart tissue was homogenized in buffer A (20 mM Tris-HCl, 1 mM magnesium acetate, 1 mM imidazole, pH 7.0, 10 mM 2-mercaptoethanol) and applied to a DEAE-Sepharose CL-6B column (Pharmacia Corp, Sweden); the eluate was applied to a CaM-Sepharose 4B column (15). Lanes A to C show the total CaM-binding protein fraction from the CaM-Sepharose 4B column eluted by 1.0 mM EGTA in buffer A in the presence of 0.2 M NaCl; 1.0 mM EGTA alone; and 1.0 mM EGTA containing 0.5 M NaCl, respectively. Lane D shows purified HMWCaMBP. Reproduced with the permission of Highwire Press from reference 15