Toxin Functionality: Human promyleocytic leukemia cell line HL60 was differentiated into neutrophils by treatment with DMSO. Neutrophils were incubated with serial dilutions of LukS and LukF at equimolar concentration for 48 hours at 37°C with 5% CO2 and 95% humidity. Cellular viability was determined by adding XTT and incubation for additional 6 hours. Cells were centrifuged and the OD determined in the supernatants at 470nm. EC50 was found to be 4.09nM of each toxin subunit.
A: SDS-PAGE analysis of 500ng of 299606. B. WB detection of LukS-PV at 50, 10, and 1ng, as well as 15ul 24-hr culture supernant of S. aureus USA300 strain. Membrane was probed with antibody 299601, followed by IgG-AP secondary Ab. The LukS from the culture medum is not His-tagged and migrates at slightly lower MW.
Supplier Page from United States Biological for PVL LukS, Recombinant, His-Tagged