Fairchild, Craig D.
HFR1 is involved in the far-red light signaling mediated by phytochrome A in Arabidopsis. In order to help elucidate the functional and regulatory role of HFR1 we used various immunodetection procedures. Immunoblotting and immunoprecipitation were employed to detect HFR1 using epitope tagged transgenic Arabidopsis, and antibodies produced to recombinant E. coli HFR1. A fusion protein was constructed and used to purify antibodies specific to the carboxy-terminus of HFR1. While efforts continue to detect HFR1 in plant extracts, the sensitivity of antibodies was determined. Purification of antibodies specific to the carboxy terminal of HFR1 was accomplished using the fusion protein. In the future, these antibodies could be used for in situ localization of HFR1. The ability to recognize HFR1 with specific antibodies in planta will allow for affinity purification and subsequent biochemical studies of HFR1.
Worcester Polytechnic Institute
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Chemistry and Biochemistry