Bacterial Toxin Research Citations

… Anthrax lethal factor (#172C) and Anthrax protective antigen (#171E) were from List Biological Labs, Inc. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

… Anthrax Protective Antigen List Biologicals Ref: 771B…

Chemicals, Peptides, and Recombinant Proteins

Protective antigen (PA), recombinant from Bacillus anthracis List Biological Laboratories 171C

Stimulation assays

… For the stimulation of the NLRC4 inflammasome, human macrophages were primed with 200 ng ml-1 LPS for 3 hours before 2 μg ml-1 LFn-PrgI and 0.5 μg ml-1 PA (from Bacillus anthracis) were added to the culture medium for 2 hours (Zhao et al., 2011).

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

Antibody quantification by ELISA

PA, LF and EF ELISAs were performed using commercially available antigens (List Biologicals, Campbell, CA) as previously described [21]. Cutaneous anthrax and regional control group sera were titered using 2-fold dilutions, beginning at 1:10. US control sera were titered at 10-fold dilutions, beginning at 1:10. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #178A – Anthrax Edema Factor (EF), Recombinant from B. anthracis

… B. anthracis lethal factor (LF) was acquired from List Biologicals. …

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

Materials and Methods

Anthrax lethal factor, Product #169 and MAPKKide® Plus, Prod #532 are from List Biological Laboratories,
Inc. The 96-well, black, flat bottom, nonbinding plates used for the fluorescent plate assay were from
Corning (cat # 3991). The filtered adult bovine plasmas: Na-Citrate, cat #7310806; K2-EDTA, cat #7310807;
K3-EDTA, cat #7310808; Li-Heparin, cat #7310810 ; Na-Heparin, cat #7310811; and the filtered adult bovine
serum cat #7330800 were purchased from Lampire Biological Laboratories.

Sample Preparation: Stock solutions of the fluorogenic substrate, MAPKKide® Plus, were made 1.25 mM
in DMSO. The substrate was then diluted 5-fold to 250 µM in assay buffer: 20 mM HEPES, pH 8.0 containing
0.1% Tween-20.

LF Activity Assay: LF was spiked into each plasma or serum sample and then each was diluted 1:5 using
assay buffer. The reaction was initiated by addition of 10 μM MAPKKide® Plus directly to the 1:5 diluted
plasma or serum, and the assays were run at 37°C using the kinetic mode of the plate reader with readings
at 1-minute intervals. The samples were run in triplicate with 9 replicate blanks. The limit of detection was
calculated from the normal distribution of the blank samples (mean + 3 stdev; n = 3 triplicates).

• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence
• Product #532 – MAPKKide® Plus (AMC) Specific Substrate for Anthrax Lethal Factor

… A total of 31.7 nM PA alone (List Labs, no. 171E) or in the presence of 13.6 nM LFN, LFN-ACD, or LFN-RID was added to the media, and cells were incubated for 20 hours at 37C in the presence of 5% CO2 …

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

Reagents

Lethal factor (LF) was purchased from List Biological Laboratories (#169, recombinant from B. anthracis)

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

Protein expression and purification

Upon addition of double molar excess of trypsin inhibitor (Sigma Aldrich), PA7 was further purified by size exclusion chromatography using a Superdex 200 column (GE Healthcare). Lyophilized LF (List Biological Lab. Inc., Lot#1692A1B) were resuspended in water according to the manufacturer’s manual and mixed with PA7 in a molar ration of 10:1. …

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

Reagents

… Lethal Toxin was from List Biological Laboratories. …

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

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

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