Fig. 2
From: Fluorescence ImmunoPrecipitation (FLIP): a Novel Assay for High-Throughput IP
a Schematic of HuEV-A expression. ColE1 ori = bacterial origin of replication; Amp. = Ampicillin resistance cassette; EBNA-1 = Epstein-Barr nuclear antigen 1; OriP = origin of plasmid replication; Gateway = Gateway cloning cassette; Tet-CMV prom. = Doxycycline inducible promoter; FRT = flippase recognition target; FLP = flippase recombinase; LoxP = Lox sequence; Cre = Cre recombinase; TEV = TEV protease cleavage site. Note that treatment with Cre recombinase or FLP recombinase can produce a “short tag” or untagged derivative of the originally cloned ORF, respectively. b Immunoblot of cell lysates from HeLa cells expressing and empty HuEV-A vector or HES1, URI, or Art-27 proteins expressed from the HuEV-A expression vector. A version of the HuEV-A expression vectors not containing YFP in the tag (after Cre treatment of the vector) was also used for each of the proteins and for the empty vector. The proteins were detected with a FLAG-M2 antibody shown in green. Tubulin (shown in red) was used as loading control