A highly sensitive assay for proteases using staphylococcal protein A fused with enhanced green fluorescent protein.

نویسندگان

  • Hiroyoshi Fujino
  • Takashi Aoki
  • Hiroyuki Watabe
چکیده

Enhanced green fluorescent protein (EGFP) was fused with staphylococcal protein A (SpA) and used as a substrate for proteases. An SpA-EGFP assay was done in three steps: (i) digestion of SpA-EGFP by proteases, (ii) addition of rabbit IgG immobilized on Sepharose beads, and (iii) measurement of the fluorescence intensity of supernatant. The assay was sensitive enough to measure picogram levels of trypsin and chymotrypsin, and may be applicable to various other proteases as one of the most sensitive methods.

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عنوان ژورنال:
  • Bioscience, biotechnology, and biochemistry

دوره 66 7  شماره 

صفحات  -

تاریخ انتشار 2002