Purification and preparation of antibody to RNA polymerase II stimulatory factors from Ehrlich ascites tumor cells.
An improved method was developed for purification of the protein termed S-II that specifically stimulates RNA polymerase II of Ehrlich ascites tumor cells. The specific activity of the final preparation was 400 000 units/mg of protein, which is about 30-fold higher than that of the previous preparation [Sekimizu, K., et al. (1976) Biochemistry 15, 5064]. The final preparation gave a single band on both sodium dodecyl sulfate and nondenaturing gel electrophoresis, and the protein extracted from the band on nondenaturing gel had stimulatory activity. S-II is a basic protein with a molecular weight of 40 500. The fundamental characteristics of S-II determined with the previous preparation were confirmed with completely purified S-II. A specific antibody to S-II was prepared. This antibody inhibited only the stimulatory activity of S-II and did not affect the activity of RNA polymerase II itself. Thus, S-II is probably not a component of the multimeric proteins of RNA polymerase II.
D000818:Animals; D000906:Antibodies; D000937:Antigen-Antibody Reactions; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004789:Enzyme Activation; D005779:Immunodiffusion; D007074:Immunoglobulin G; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000502:physiology*; D012319:RNA Polymerase II / Q000302:isolation & purification / Q000378:metabolism*
D000906:Antibodies; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases