Native species of helix destabilizing protein-1 in mouse myeloma identified by antibody probing of Western blots.
Antibody probing of Western blots is a method for analyzing the apparent Mr of a protein in any given preparation (Renart, J., Reiser, J., and Stark, G., Proc. Natl. Acad. Sci. USA 76: 3116, 1979). We prepared a rabbit antiserum to purified mouse myeloma helix destabilizing protein-1 and used this antiserum in Western blotting experiments with a crude homogenate of mouse myeloma. The results indicated that the native species of helix destabilizing protein-1 can be degraded during purification. This in vitro proteolysis results in complete loss of the native species and accumulation of lower Mr proteins that represent limit digestion products. These findings have identified the true native form of mouse myeloma HDP-1 as a protein of apparent Mr = 36,000 to 38,000, instead of the Mr = 24,000 and 27,000 proteins obtained by routine purification.
D000818:Animals; D003101:Collodion; D004265:DNA Helicases / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008970:Molecular Weight; D009101:Multiple Myeloma / Q000032:analysis*; D010447:Peptide Hydrolases / Q000378:metabolism; D011817:Rabbits / Q000276:immunology; D012261:Ribonucleoproteins; D013951:Thymus Hormones; D014764:Viral Proteins
D004268:DNA-Binding Proteins; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; C000619657:Hnrnpa1 protein, mouse; D007106:Immune Sera; D012261:Ribonucleoproteins; D013951:Thymus Hormones; D014764:Viral Proteins; C060035:gp32 protein, Enterobacteria phage T4; C027729:helix-destabilizing proteins; D003101:Collodion; D010447:Peptide Hydrolases; D004265:DNA Helicases