Polyclonal antibodies against rat liver cytosolic casein kinase II (CK-2) cross-react with CK-2 from other tissues and nuclear form (PK-N2) of the enzyme.
Casein kinase II (CK-2) is a ubiquitous serine/threonine protein kinase, and is localized to both the cell nucleus and cytoplasm. Despite extensive biochemical similarities in their properties, there is evidence that the two forms of the enzyme exhibit certain distinctions (1). This prompted us to produce antibodies against CK-2, which could be utilized as a possible tool for investigations of the various forms of this enzyme. Specific polyclonal antibodies against the rat liver cytosolic CK-2 were raised in egg yolk of laying hens; the enzyme had repeatedly failed to elicit an immunogenic response in rabbits. The purified polyclonal antibody (egg yolk immunoglobulin, IgY) recognized all three subunits (42, 38, and 28 kDa) of the enzyme in immunoblots. The antibody when bound to a matrix was capable of removing CK-2 from solution, and the bound enzyme could be recovered from the immunoaffinity matrix with 0.1 M diethylamine. The antibody exhibited a high affinity towards CK-2 prepared from cytosol of liver, ventral prostate, and several other rat tissues, but no immunoreactivity was detected towards a number of other protein kinases tested. The subunits of the nuclear form of CK-2 (PK-N2) migrated differently when electrophoresed in parallel in the same gel. However, the antibody did cross-react with the various subunits of PK-N2 suggesting a significant homology in the immunogenic domains in the various subunits of the two forms of the enzyme.
Goueli
S A
SA
Department of Laboratory Medicine and Pathology, University of Minnesota School of Medicine, Minneapolis.
Hanten
J
J
Davis
A
A
Ahmed
K
K
D000818:Animals; D000906:Antibodies; D047388:Casein Kinases; D002467:Cell Nucleus / Q000201:enzymology; D003429:Cross Reactions; D003600:Cytosol / Q000201:enzymology; D007120:Immunochemistry; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D051381:Rats; D014018:Tissue Distribution
D000906:Antibodies; D011494:Protein Kinases; D047388:Casein Kinases