E. coli Protein Expression with HiBiT Quantification

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E. coli Protein Expression with HiBiT Quantification is a fully automated end-to-end workflow for expressing HiBiT-tagged proteins in E. coli and quantifying expression via lumines cence. Beginning with bacterial heat shock transformation in 384-well format, the protocol proceeds through glycerol-based culture inoculation, 16-hour outgrowth, cell pelleting, mechanical lysis with shaking incubation, and luminescence detection using the BMG PHERAstar plate reader. Key automation instruments include Inheco shakers/incubators, Cytomat growth chambers, Agilent Bravo 96 and 384 liquid handlers, Labcyte Echo acoustic dispenser, BioTek MultiFlo dispenser, and BMG PHERAstar. Results are reported as raw and standard-curve-normalized luminescence values (bcRLU), enabling quantitative comparison of HiBiT-tagged protein expression across up to 384 constructs per run. Show more