A. Mainero et al., RABBIT CERULOPLASMIN - PURIFICATION AND PARTIAL CHARACTERIZATION, Preparative biochemistry & biotechnology, 26(3-4), 1996, pp. 217-228
Rabbit ceruloplasmin (Cp) was purified after solid ammonium sulfate pr
ecipitation to 60% final saturation by a two-step column chromatograph
y procedure, utilizing DEAE-Sephadex A-50 and changing the NaCl concen
tration in the buffer to 0.16 M to achieve the isolation of the protei
n. The purified Cp was used to prepare antibodies in guinea pigs that
were used afterwards to determine Cp concentration in normal rabbits a
nd in rabbits with an experimentally induced chronic anemia. The molec
ular,weight of rabbit Cp determined by SDS-PAGE was 125,000 and a high
molecular weight Cp of 200,000 comprising 8% of the total purified pr
otein was also found. An optical density ratio (610 nm/280 nm) of 0.04
75 and a molar extinction coefficient of 7625 were obtained. Copper de
terminations yielded a value of 0.24% that corresponds to 5 copper ato
ms per molecule. The staining of Cp following disc-electrophoresis in
polyacrylamide gels also showed a two band pattern.