Citations

Materials and reagents were obtained from … LF, PA83, and PA63 were obtained from List Biological Laboratories (Campbell, CA), …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Reagents

… Bacillus anthracis PA was obtained from List Biological Laboratories. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

… A 10× solution of the lethal factor (LF, List Biological Labs) in medium was prepared at 5 µg/mL, and 10 µL of the 10× stock was added to the well to achieve a 0.5 µg/mL …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Immunoblotting and pull-down assays

Cell lysates and cell culture supernatants were separated by 10% or 15% SDS-polyacrylamide gel electrophoresis under reducing conditions (non-reducing when using the antibody against C5), transferred to a nitrocellulose membrane and blocked with 5% non-fat dry milk in 0.1% Tween 20/TBS solution. The membranes were probed with primary goat antibodies against PA (1:5000, List Biological Laboratories), …

… The aLF was ordered from List Biological Laboratories, Inc (Listlabs, US). …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… Goat anti-PA (B. Anthracis) List Biological Laboratories Cat# 771B …

2.1 Materials and Chemicals

… The aLF was ordered from List Biological Laboratories, Inc (Listlabs, US). …

2.2 The Preparation of aLF Proteolytic Reaction

aLF, ZnCl2 and peptide substrate were mixed together as proteolytic buffer (5.64 μM, 28 mM and 2.82 μM) for endopeptidase activity reaction, …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… Recombinant protective antigen and edema factor were obtained from List Biologicals (Campbell, CA, USA) …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #178A – Anthrax Edema Factor (EF), Recombinant from B. anthracis

4.4. ELISA

Ninety-six-well microtiter plates (Nunc Maxisorp, Sigma Aldrich, L’lsle-d’Abeau Chesne, France) were coated by incubation with PA83, LF (List laboratories) or KLH (as control; Sigma Aldrich, L’lsle-d’Abeau Chesne, France) diluted in PBS (5 µg/mL, 100 µL per well) overnight at 4 °C. …

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

Anti-Protective Antigen (PA) Enzyme-Linked Immunosorbent Assay

In this study, serum samples were assessed for the presence of specific antibodies against the anthrax PA as described by Yu et al. (56), and Ndumnego et al. (23). Briefly, microtiter plates (Thermo Scientific™ Pierce 96-well Plates-Corner, USA) were coated overnight with 0.5 μg/ml rPA (List Biological Laboratories Inc., USA) in bicarbonate buffer at 4°C. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis