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Screening libraries with protein probes
Expression and purification of recombinant antigens
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activation-tagged cDNA affinity chromatography agar agarose amino acids ampicillin antigen assay baculovirus bait beads binding blot cDNA cDNA encoded proteins centrifugation cloning coli colonies column containing described in Protocol dilution EDTA efficiency elution enzyme epitopes Equipment and reagents expression system expression vectors Figure filter foreign gene foreign protein fusion protein Gal/Raf galactose genome Glu ura glucose glutathione hybridoma inclusion bodies incubate induced insect cells insert insoluble lacZ lacZ reporter lane Leu+ LEU2 LexA library plasmid lncubate medium mg/ml microcentrifuge molecular monoclonal antibodies NaCl overnight peptide pGEX plaques plasmid polyclonal antibodies preparation production protease protein expression Protocol 9 purification recombinant protein resin Resuspend the pellet room temperature sample screening SDS-PAGE secreted secretion vectors selection sequences sodium soluble solution sterile strain supernatant target protein transcription transformants Tris-HCl pH tube virus Wash Western blot X-Gal X-Gal plates yeast