Bacterial Toxin Research Citations

STAR★Methods
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Chemicals, peptides, and recombinant proteins

Lethal factor List Biologicals Cat#172B

Protective antigen List Biologicals Cat#171E

Activation of BMDMs and human macrophages

All cells were grown, maintained and activated at 37°C and 5% CO2 in a water-jacketed incubator. 1 × 106 WT or Casp1/11 DKO murine macrophages were plated per 12-well in DMEM, 10% (v/v) FCS (DMEM complete medium) one day prior to stimulation. All BMDMs were primed with 200 ng/mL LPS from Salmonella Typhimurium for 2 h or left untreated and subsequently washed three times with serum- and phenol red-free DMEM supplemented with 2 mM L-glutamine (DMEM minimal medium). For the time course experiment (Dataset 1), cells were preincubated with 0 or 100 μM glyburide (#NBP2-30141, Novus Biologicals) for 15-30min before 2 mM ATP (#7699, Sigma) or 10 μM nigericin (N1495, Invitrogen) were added for 15, 30, 45, and 60 min. Lethal factor and protective antigen were added at 1ug/ml. …

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

ELISA

The three strains cultured in TY medium for 12, 24, 48, and 72 h were centrifuged to collect the supernatants containing TcdA and TcdB toxins; the supernatant was filtered with a 0.45 µm filter. Next, a 96-well plate was coated with the supernatant at 4 °C overnight and then washed with PBST (phosphate-buffered saline pH 7.4 + 0.05% Tween 20). The plate was blocked with 5% skim milk at 37 °C for 2 h. The polyclonal antibodies of TcdA and TcdB (chicken IgY; 1:1000 dilution; List Biological laboratories) were used for first hybridization; then, goat anti-chicken IgY secondary antibody, (HRP-conjugated; 1:5000 dilution; Invitrogen) was used. The TMB chromogenic reagent kit (Sangon, China) was used to determine the absorbance at 450 nm. Pure toxin A and toxin B (List Biological laboratories) were used as positive controls, and the standard curves were obtained using 20, 2, 0.2, 0.02, and 0.002 ng/ml of TcdA and TcdB [19].

• Product #152C – Toxin A from Clostridium difficile
• Product #155 – Toxin B from Clostridium difficile

Induction and assessment of EAE

C57BL/6 mice were immunized by subcutaneous injection with 150 μg of MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK, Sigma-Aldrich) emulsified in a complete Freund’s adjuvant containing 5 mg/ml H37RA (Difco Laboratories). On day 0 and 2 after immunization, 200 ng pertussis toxin (List Biological Laboratories) was given intravenously. …

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)

EAE Induction and Treatment

EAE was induced in 8-week-old mice by subcutaneous immunization with 150 μg MOG35–55 peptide emulsified in complete Freund adjuvant (Difco Laboratories, Detroit,
MI) per mouse, followed by administration of 200 ng pertussis toxin (List biological Laboratories, Inc., Campbell, CA) on days 0 and 2 as described. …

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)

… In each of three locations, 100 µl volume was injected s.c. Pertussis toxin (200 ng in 100 µl PBS; List Biological Laboratories) was injected i.p. on the day of immunization …

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)

Diabetes Induction

Homozygous PANIC-ATTAC mice were administered a single dose of AP20187 homodimerizer (0.08125 mg/kg) prepared according to the manufacturer’s instructions (17). Female NOD mice were surveyed twice-weekly by glucometer, until spontaneous diabetes was diagnosed by glucose >250 mg/dL for two consecutive measurements. DT (List Biological Laboratories; 150) was administered to NSG-DTR mice in a single, 300-μL intraperitoneal injection of 5.0 ng total DT. Control NSG-DTR mice (phosphate-buffered saline; PBS) were treated with an equal volume of 1× PBS (Sigma-Aldrich). All animals in a cohort (with human islets from the same human donor) were injected with DT or PBS on the same day (29).

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

… Diphtheria toxoid (60 µg) (List Biological Laboratories, Campbell, CA) and a negative control of recombinant Schistosoma mansoni glutathione-S-transferase (GST) (15 µg) (Moss et al., 2004) were each coupled to 1.25 × 10 7 beads in a 1 ml volume of buffer containing …

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

… Adjuvanted formulations contained cholera toxin B (List Biological, cat #104) …

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

… Pertussis toxin List Biological Laboratories Cat# 180…

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

… Then ultrapure LPS (0.01 µg/mL, List Laboratories, Campbell, CA) was added to the wells in the continued presence or absence of ERK1/2 inhibitor (1 µM) …

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/