Production of a polyclonal antibody raised against recombinant flounder p53 protein.
The cDNA encoding the wild type p53 protein from flounder, Platichthys flesus, was expressed in Escherichia coli using the GST fusion protein system. Several milligrams of recombinant p53 protein were purified. This protein displayed an apparent molecular weight of 45,000 Da, a value which is very similar to Xenopus p53, but significantly greater than was expected based on the length of the open reading frame. Immunization of rabbits against purified p53 protein allowed the production of high titre polyclonal antiserum. This new polyclonal antibody recognized recombinant flounder p53 protein in Western blot. Cross reaction was also observed with recombinant Xenopus p53 protein but not with human p53 protein. Immunoblotting of the total protein extract from normal flounder ovaries did not reveal any p53 expression.
Cachot
J
J
Laboratory of Ecotoxicology, IFREMER, Nantes, France. jcachot@ifremer.fr
Galgani
F
F
Vincent
F
F
D000818:Animals; D000906:Antibodies / Q000032:analysis; D000918:Antibody Specificity; D015153:Blotting, Western; D003429:Cross Reactions; D018076:DNA, Complementary / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005432:Flounder; D005978:Glutathione / Q000378:metabolism; D007106:Immune Sera / Q000096:biosynthesis* / Q000276:immunology; D008970:Molecular Weight; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D018076:DNA, Complementary; D007106:Immune Sera; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; D005978:Glutathione