ELISPOT:
Peripheral blood mononuclear cells (PBMCs) were prepared and washed in RPMI medium supplemented with 10% fetal calf serum, penicillin, streptomycin, and L-glutamine. …The antigen, recombinant Protective Antigen (rPA), LF or EF (List Biological Laboratories Campbell, Calif.) was added at a final concentration of 12.5 g/ml for PA or 25 g/ml for either LF or EF. The plates were incubated overnight at 37C in a 5% CO2, humidified atmosphere. …
Quantitative anti-PA IgG ELISA:
Wells of 96-well microtiter plates (Immulon 2HB; Dynex Technologies, Chantilly, Va.) were coated by adding of 100 l rPA (List Biological Laboratories Campbell, Calif.) diluted to 1 g/ml in PBS. Plates were incubated overnight at 4C. …
Lethal toxin neutralization assay (TNA):
…Lethal toxin (LeTx; 100 ng of rPA/ml and 50 ng of LF/ml, final concentrations) was prepared in the same medium and pre-incubated with the sample dilutions in a humidified incubator set at 37C, 5% CO2 for 1 h. LeTx was prepared using PA obtained from List Biological Laboratories …
End-point ELISA for LF and EF:
Wells of 96-well microtitre plates (Immulon 2HB) were coated with either LF or EF (List Biological Laboratories) at 1 g/ mL in PBS (100 L per well). …
Author did not indicate which specific lethal factor was utilized. List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).
Author did not specify which List Labs Anthrax Edema Factor was utilized. List Labs provides Product #178A(Anthrax Edema Factor (EF), Recombinant from B. anthracis) and Product #173B (Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis).