Bacterial Toxin Research Citations

… Anti-PA IgG was measured in rabbit sera via electrochemiluminescence similar to the PA-ECL screening assay. Biotinylated recombinant PA83 (List Biological Laboratories, Inc.) was bound to streptavidin-coated plates (MSD) and used as the capture antigen. …

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

NLRC4 Activity in Human Monocytes

A 10-µL aliquot of compound (10X in 1% DMSO) was added to PMA-differentiated THP1-Null2 cells (65,000 cells in 80 µL of RPMI media per well) and incubated for 60 minutes in a humidified incubator at 37°C with 5% CO2. NLRC4 activation was initiated by adding a 10 µL mixture of LFnNeedle and B. anthracis protective antigen (PA) (List Labs #171E) (10X). LFn-Needle is fused to the amino-terminal domain of LFn (in house). This fusion system, when co-administered with the anthrax toxin’s PA, allows intracellular delivery of the bacterial ligand [11]. OptiMEM (10 µL) was added to control wells in lieu of LFnNeedle/PA treatment. The plate was incubated for 5 hours in a humidified incubator at 37°C with 5% CO2.

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

2.13. Priming of Inflammasomes

Inflammasome activation was measured according to Primiano et al.’s description [28]. HCM3 cells were plated in EMEM without supplements at 1 × 105 cells/well. Cells were stimulated for 1 h with 10 g/mL (13.4 mM) of nigericin (tlrl-nig, InvivoGen, San Diego, CA, USA) for 2 h with 2 mM ATP (tlrl-atpl, InvivoGen), or overnight with 200 g/mL of monosodium urate crystals (MSU; tlrl-msu, InvivoGen) for NLRP3 induction; flagellin/DOTAP complex, obtained with 25 μL HBS buffer with 500 ng flagellin (final concentration at 1 μg/mL; tlrl-stfla, InvivoGen) and 3 μL DOTAP (11202375001, Sigma-Aldrich) for each well for NLRC4 induction; with 0.5 mL Opti-MEM supplemented with GlutaMax (Opti-MEM™ Reduced Serum Medium containing GlutaMAX™ Supplement, 51985034, Gibco, Grand Island, NY, USA) containing 1 μg/mL Bacillus anthracis lethal factor (172B, List Biological Laboratories, Campbell, CA, USA) for 3–6 h for NLRP1 induction; and with a poly(dA:dT)/lipofectamine complex, obtained by mixing 50 μL Opti-MEM containing 1 μg poly(dA:dT) (final concentration at 2 μg/mL) with 50 μL Opti-MEM containing 2.5 μL lipofectamine 2000 (11668027, Thermo Fisher’s Scientific, Waltham, MA, USA) for each well for AIM2 induction. …

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

Anthrax lethal toxin neutralization assay

RAW264.7 cells were seeded as mentioned above and cultured in DMEM supplemented with 10% FBS and 10 mM HEPES (Fisher Scientific, Fai Lawn, NJ, USA) overnight. The toxin neutralization assay was performed as previously described [73]. Pooled serum samples from vaccinated guinea pigs before challenging with Bcbva UFBc0009 and B. anthracis Ames UF00738 spores were prepared at 200-, 1,200-, and 12,000-fold dilutions in DMEM supplemented with 10% FBS and 10 mM HEPES. Pooled serum samples from unvaccinated animals served as a control. The diluted serum samples were then incubated with 50 ng/ml PA (List Labs, Campbell, CA, USA) and 160 ng/ml LF (List Labs, Campbell, CA, USA) for 30 min before adding to RAW264.7 cells. …

• 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

Stimulations and viral infections

N/TERT1 cells were plated at 5 × 104 cells/96-well plate for cytokine release experiments and at 2.5 × 105, 5 × 105, or at 1 × 106 cells per 24-, 12-, or 6-well plate, respectively. The next day, the cells were treated as indicated with 2 µM Val-boroPro (ApexBio and Biozol), 2 µM ANS (Biomol), 100 ng/mL TNFα (PeproTech), 6.5 µM Nigericin (Sigma Aldrich), 0.1 µM B/B homodimerizer (AP20187, MedChem Express), 20 µM Z-VAD-FMK (Peptanova), 10 µM cytarabine (Ara C, Sigma Aldrich), 1 µM MLN4924 (MLN), 1 µg/mL anthrax lethal factor, and 1 µg/mL protective antigen (both List Biological Labs). …

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

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

Chemicals and reagents

… LF (List Labs), …

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

Immunogenicity Studies of PA83 variants in Mice

Immunogenicity studies were performed with 3 and 10 μg of plant-produced dPA83(E), dPA83(P) and commercial recombinant PA (rPA, List Biological Laboratories, Campbell, CA, used as a positive control) proteins, administered intraperitoneally in seven- to eight-week-old Balb/c mice (6 animals/group). Mice were injected with three doses of protein samples adsorbed in 0.3% Alhydrogel at three-week intervals (day 0, 21 and 42). …

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

… Anthrax lethal toxin List Biological Laboratories Inc. #171B/#172B …

Treatments

See Table S1 for list of Figures/panels in each figure and corresponding dose and exposure time. General information for the methods is provided here. To induce cytokine release from macrophages, microglia and BMDM cells were primed for 4–5 h (or time indicated in Table S1)with 1 μg/mL of LPS in culture medium, and some cells were pre-treated with 1 μM of CP456773/MCC950 for 30 min before ATP stimulation. Cells were then pulsed with 5 mM ATP for 30 min. For cytokine induction by anthrax lethal toxin (LeTx), LPS-primed cells were treated with LeTx at a final concentration of 250 ng/mL (LF and PA) for 3 h. Cell supernatants were collected and frozen at −80°C until assayed.

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

… goat anti-EF (773L, List Labs, Campbell, CA, USA), …