Following in vitro transcription and translation of JV V5 a new product, approximately 42 kDa in size, was visible ( Figure 5 , lane 2). This product was not observed in any prior analyses of JV. To confirm that this protein was V5/N-term associated the TNTH reaction was subjected to RIPA using the anti-V5 antibody ( Figure 5, lane 3) . This confirmed expression of the N-term protein in vitro.