Sensitive Detection of Rodent Rb Protein by Polyclonal Antibodies Against a Bacterially Expressed Mouse Rb Protein

抄録

Polyclonal antibodies against the bacterially expressed mouse retinoblastoma gene product (pRb) were prepared and characterized to enable sensitive recognition of rodent pRb. The cDNA fragment corresponding to amino acids 237-595 of the mouse pRb sequence was expressed at a high level in E. coli as a fusion protein with glutathione S-transferase. Polyclonal anti-mouse pRb antibodies, MKS-1, were prepared by immunizing rabbits with the truncated pRb obtained from the fusion protein. MKS-1 sensitively recognized the hyper- and the hypophosphorylated pRbs not only in the rodent cells but also in primate cells by Western blotting following immunoprecipitation. An increase in the cellular content of total and hypophosphorylated pRbs during myogenic differentiation of mouse C2 cells was also detected quantitatively by MKS-1. Thus, MKS-1 is useful for the quantitative detection and the analysis of pRb in cell lines established from rodent tissues.