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. 2002 Oct 1;30(19):4166–4175. doi: 10.1093/nar/gkf548

Figure 5.

Figure 5

Immunodetection of H.volcanii SRP54. (A) Aliquots of wild-type E.coli cells, IPTG-induced E.coli cells transformed with plasmid pET-Hv54 encoding for H.volcanii SRP54, or Ni–NTA-purified H.volcanii SRP54 were probed with polyclonal antiserum raised against the H.volcanii SRP54. The specificity of antibody labeling of the bacterially expressed archaeal protein was confirmed by the failure of normal rabbit serum (NRS) to label such a band in E.coli cell extracts. Similarly, the polyclonal antibodies recognized SRP54 in an extract of H.volcanii. Again, NRS failed to label any such band. (B) Aliquots of H.volcanii cells, isolated membranes, the S100 cytoplasmic fraction and isolated ribosomes were probed with anti-H.volcanii SRP54 antibodies. The S100 and ribosomal fractions were prepared as described by Ban et al. (39). In both (A) and (B), the arrow depicts the position of the labeled 54 kDa protein recognized by the antibodies, while molecular weight markers shown on the right denote the 250, 150, 100, 75, 50, 37 and 25 kDa positions.