Purification of a fragment of MDM2 for production of polyclonal antisera.
A histidine-tagged, carboxy-terminal fragment of the murine double minute 2 gene product, p90(MDM2), was purified by Ni--NTA chromatography and preparative gel electrophoresis. The purified MDM2 fragment was used to generate polyclonal antisera that recognize multiple species of MDM2 proteins, including the inhibitor of p53, p90(MDM2), as well as the activator of p53, p76(MDM2). The antibodies are useful for Western analysis, immunoprecipitation, and immunofluorescence.
Department of Oncology, McArdle Laboratory for Cancer Research, Madison, Wisconsin 53706, USA.
B P
BP
Carstens
M E
ME
Perry
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004926:Escherichia coli; D005455:Fluorescent Antibody Technique; D006639:Histidine; D007106:Immune Sera / Q000096:biosynthesis / Q000276:immunology*; D051379:Mice; D009532:Nickel / Q000378:metabolism; D009687:Nuclear Proteins; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D011233:Precipitin Tests; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism
D007106:Immune Sera; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010455:Peptides; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; C033223:polyhistidine; D006639:Histidine; D009532:Nickel; C497207:Mdm2 protein, mouse; D051736:Proto-Oncogene Proteins c-mdm2