Citations

Stimulation conditions:

iMos were primed with 200ng/mL LPS (Ultrapure LPS, Invivogen) for 3h. Inflammasome activation was performed with 10M nigericin (Life Technologies) and 5mM ATP (Sigma) for 1h or with 1mg/mL Silica (US-Silica), 1mM LeuLeuoMe (ChemImpex), lethal toxin (1g/mL lethal factor (List Biologicals), 1g/mL, protective antigen (List Biologicals), poly(dA:dT) (20ng/mL+0.5% lipofectamin 2000), LFn-PrgI (4g/mL (M. Geyer)+protective antigen 1g/mL) for 3 to 4h, 12 or 25g/mL PGN from S. Aureus (Invivogen) for 20h, 20g/mL R837 (Invivogen) for 1.5h, cytosolic LPS (Cholera toxin subunit B (CTB) (20g/mL+2g/mL LPS), or with DOTAP (3.75g/mL+750ng/mL LPS)) for 16h. …

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

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

… collected by gentle pipetting, centrifuged at 300 3 g for 5 min at 25C, and resuspended in 3 ml of complete RPMI 1640 medium supplemented with 10 ng/ml recombinant murine GM-CSF, 1 mg/ml LPS (LPS from Escherichia coli J5(Rc); List Biological Laboratories …

• Product #301 – LPS from Escherichia coli J5 (Rc)

… Diptheria toxin List Biological Laboratories #150 …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Muscle afferents tracing:

 CTB (1%; 10 µl, List Biologicals) was injected into the extensor diaminobenzidine (DAB; Sigma) or streptavidin conjugated to FITC or Cy3. The following primary antibodies were used: goat anti- CTB (1: 1000, List Biologicals), …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… In LPS-neutralizing assays described below, we used smooth-type LPS from Escherichia coli O55:B5 (List Biological Laboratories, CA, USA) as an endotoxin. …

• Product #203A – LPS from Escherichia coli O55:B5

… A short paralytic half-time in the MPN assay reflects a high potency of the investigated toxin. After equilibrating the tissue in EBSS for 15 minutes the bath solution was replaced by a BoNT/A (List Laboratories, Campbell, CA, USA) containing solution …

Cell Ablation:

For ablation of neurons expressing Cre and FlpO drivers in addition to Tau^ds-DTR, mice were injected with diphtheria toxin (DT; 50 ng kg^-1 in 0.9% sterile saline, i.p.; List Biological Laboratories) at P28 and P31 (Bourane et al., 2015a). Analysis of spontaneous scratching in mice expressing NPY::Cre (and relevant controls) was performed at P35, 7 days following the first DT injection. For the double-Cre ablation experiment, controls were positive for Cre and Tau^ds-DTR alleles but lacked the Lbx1^FlpO allele and did not therefore express the diphtheria toxin receptor; however, all mice received DT. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Immunohistochemistry:

The following primary antibodies were used in this study: …  goat -CTB (1:4000; List Laboratories), …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Diphtheria toxin List Biological Laboratories, Inc Un-nicked, Lot #: 15043A1 …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

… Nicked BoNT/E3 was obtained from List Biological Laboratories (Campbell, CA, USA). …

Sandwich Assay Procedure:

When performing the sandwich assay, for every mAb, the unlabeled mAb was used as the capture antibody and the biotinylated form was used as the detector antibody. Each mAb was tested in combination with all of the mAbs to identify pairs that can detect the BoNT/E3 toxin in the solution phase. Black plates were coated with 100 L of mAb at 2 g/mL in 0.05 M sodium carbonate buffer, pH 9.6, incubated overnight at 4 C, and then blocked with 3% NFDM-TBST. The blocking solution was removed and BoNT/E3 toxin in 3% NFDM-TBST was added at an initial concentration of 500 ng/mL and serially diluted two-fold, including wells with no toxin. Next, biotinylated-mAb was added in triplicate at 1 g/mL, 100 µL/well in 3% NFDM-TBST. Plates were incubated for 1 h at 37°C, washed 6 with TBST, and visualized using SuperSignal West Dura Extended Duration Substrate as described above.

When the most sensitive combination of capture mAb and detector mAb-biotin were selected, the final sandwich assays were performed in triplicate using BoNT/E3 holotoxin at a starting concentration of 1000 ng/mL, serially diluted three-fold, including wells with no toxin, and visualized using SuperSignal Femto Max Sensitivity substrate. Limit of detection cut-off values were determined using three times the standard deviation of the wells with no toxin.

EAE induction and clinical scoring:

… Pertussis toxin (500ng/mouse, List Biological Laboratories, Inc., Campbell, CA) was injected intraperitoneally (i.p.) on days 0 and 2. Animals were monitored daily for EAE signs as described⁵⁶.

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)