The Cell Biolabs Bacterial Protein Extraction Reagents contain a gentle, nonionic detergent formulation which quickly extracts functional, recombinant protein from E. coli without mechanical disruption.
Extraction of His-RFP fusion protein. His-RFP expression in BL21 was induced with 1 mM IPTG at 37ºC for 4 hrs. Cells were pelleted (left), then resuspended in Bacterial Protein Extraction Reagent for 15 minutes, according to the assay protocol. The supernatant (right) containing the extracted His-RFP was collected from the insoluble post-extraction pellet (center).
Comparison of Sonication with Bacterial Protein Extraction Reagent. Pelleted His-RFP cells were sonicated in PBS/Triton X-100 or resuspended in Bacterial Protein Extraction Reagent for 15 minutes. Lysates were then collected and tested for RFP fluorescence on a 96-well fluorometer.
Lee, J. et al. (2018). Structural Characteristics of Pneumolysin and Its Domains in a Biomimetic Solution. ACS Omega. 3(8):9453-9461. doi: 10.1021/acsomega.8b01212 (#AKR-181).