Citations

Reagents

… Lipopolysaccharide (E. coli, serotype O111:B4 L2630) was from List Biological Laboratories (Campbell, CA) …

Alcohol Exposure and LPS Challenge

Mice were exposed to ethanol (2 or 4 g/kg), propanol (2 g/kg), isopropanol (2 g/kg) or PBS via intraperitoneal injection. One hour later, a lethal dose of LPS (10 mg/kg, i.p.) was injected. Animals were monitored every 2 h for 36 h post LPS injection.

• Product #201 – LPS from Escherichia coli O111:B4

Immunization

Eight micrograms of OVA (Sigma) and 2 µg of cholera toxin subunit B (List Biological) in 6 µL of sterile PBS or 6 µL of PBS were administered intranasally. Mice were sacrificed 7 days later and parotid glands were removed and frozen …

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

Chemicals and cell culture

… cholera toxin (10 μg/ml; List Biological Laboratories, Inc., Campbell, CA, USA), …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

… All mice then received a single intraperitoneal injection of pertussis toxin (List Biological Laboratories, #180) at a concentration of 2 ng/l in 200 l PBS …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

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

Exposures

Mice received ultrapure E. coli O111:B4 LPS (List Biological Labs, Campbell, CA) (50 ml of 1 mg/ml in
PBS), or control PBS vehicle only, by oropharyngeal aspiration and were phenotyped 24 hr later. For
systemic LPS exposure, mice received 10 mg/kg LPS or PBS by intraperitoneal injection and were
phenotyped 24 hr later. …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

Breakdown of the BBB and the passive transfer of IgG to mice

After completing one session of EEG recording under normal conditions, the mice received IgG by passive transfer. Before IgG transfer, the blood–brain barrier was disrupted using a previously described method [35]. In brief, 50 μg complete Freund’s adjuvant (CFA, Sigma-Aldrich, USA) containing heat-killed H37Ra Mycobacterium tuberculosis (Difco, USA) in 50 μl was subcutaneously injected into each of four sites on the hind flank. In addition, mice received an intraperitoneal (i.p.) injection of 200 ng pertussis toxin (PTx, List Biological Laboratories, USA) in 0.2 ml PBS. The injection of PTx was repeated again 3 days later. On the 7th day after the injection of CFA, animals were injected intravenously with PBS (as vehicle) or test or control IgG (200 μg in 0.2 ml PBS). Electrophysiological recording was conducted again 1, 24, 48, and 72 h after the injection of IgG.

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)

Rat treatment, CSF and blood collection, leukocyte count

Eight-day-old (P8) rats were injected intraperitoneally (i.p.) with 1 or 5 mg/kg P3C (Invivogen, France), 0.3 mg/kg ultrapure LPS (List Biological Laboratory, USA), or 0.9% saline. …

Author did not specify which List Labs LPS product was utilized in their research.
List Labs provides the following LPS products: https://listlabs.com/product-information/lipopolysaccharides/

… emulsified in complete Freund’s adjuvant (CFA; 1.5mg/ml) followed by inoculations with 300ng of pertussis toxin, Bordatella pertussis, (List Biological Labs) on the day of induction and 48 hours later. …

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)

… LPS was purchased from List Biological Laboratories, Inc. …

Author did not specify which List Labs LPS product was utilized in their research.
List Labs provides the following LPS products: https://listlabs.com/product-information/lipopolysaccharides/