Citations

Injection of a retrograde tracer into the parafacial respiratory group

In five animals, pFRG injections of the retrograde tracer cholera toxin b (CTb, 1% in deionized water; List Biological, Campbell, CA) were unilaterally performed. For pFRG injections, the dorsal surface of the brain was exposed via an occipital craniotomy. The stereotaxic coordinates for targeting the pFRG were 2.7 mm caudal from lambda, 2.6 mm lateral from midline, and 8.7 mm ventral from the skull surface. The CTb (30 nl) was delivered by pressure through glass pipettes. …

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

… CTb Gt Polyclonal List Biological Labs 102946 502 7032A10 1:5000 1:10000 …

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

C. difficile in vitro Vero cell cytotoxicity assay

… The cytotoxic titer was defined as the reciprocal of the highest dilution that produced rounding in 80% of Vero cells for each sample. Vero cells treated with purified C. difficile toxins (A and B) and antitoxin (List Biological Labs, 152C and 155C; TechLabs, T5000) were used as controls.

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

Key Resources Table

LPS ultra pure from Salmonella minnesota R595 List Biological Laboratories, INC. Cat# 434

Infections and stimulations

MDDCs were infected or stimulated with innate agonists between day 4 and day 6 after differentiation. For infections with HIV-1 and related lentiviruses, except where noted otherwise, Vpx+ virus-like particles were used to overcome virus restriction (1 mL supernatant for 1 × 10ˆ7 cells) (Johnson et al., 2018, Manel et al., 2010). MDDCs were counted on day 4 and resuspended at 800,000 cell per ml in fresh medium with GM-CSF, IL-4, and polybrene (1 μg/ml) and then reseeded into appropriate culture vessels. For most assays, MDDCs were infected at a density of 800,000 cells per ml by diluting virus in MDDC media (without cytokines or polybrene) to a final volume normalized to controls. Innate and inflammatory stimuli were used at the indicated concentrations: poly(I:C) (InvivoGen, 10 μg/ml); LPS (List Biological Laboratories, INC, 1 ng/ml).

• Product #434 – ULTRA PURE LPS from Salmonella minnesota R595 (Re)

… Pertussis toxin (List Biological) (400 ng) was injected on the day of the immunization and again two days 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)

Antibody selection in microtiter plate

The antibody selection was performed as described previously with modifications33. In brief, for the panning procedure, the antigen was immobilized on a Costar Highbinding 96 well plate (Corning, New York, USA). 1 µg of DT (List Biological Laboratories, Campbell, USA) was diluted in PBS (137 mM NaCl; 1.76 mM KH2PO4 × 2 H2O) and coated in the wells at 4 °C overnight. …

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

Antibody ELISA assay. BPZE1 and BP611-98 bacterial lysates were prepared as described in the Supplemental Materials and Methods. FHA, PT, PRN, FIM2/3, and ACT were purchased from List Biological Laboratories, USA …

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)

• Product #186 – Fimbriae 2/3 from B. pertussis
• Product #187 – Pertactin from B. pertussis (69 kDa Protein)
• Product #188L – Adenylate Cyclase Toxin, Recombinant from B. pertussis

… At the same time and 3 d later, pertussis toxin (300 ng pertussis toxin in PBS; List Biological) was ip injected. On day 21 after MOG35-55 injection, B cells were sorted by B220 MicroBeads. …

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)

Analysis of axonal regeneration following SCI.

Adult 8-week-old cre+Pfn1 mice and 15-week-old female C57BL/6 mice were deeply anesthetized with isoflurane. Laminectomy was performed at the thoracic T8 to T9 level and the spinal cord was cut using a micro feather ophthalmic scalpel (Feather, Safety Razor Co). For CL experiments, animals were subjected to a sciatic nerve transection 1 week prior to SCI. In cre+Pfn1 mice, analysis of dorsal column axon regeneration after either SCI or CL was performed 4 weeks after injury. Dorsal column axons were traced by injecting 2 μL of 1% CT-B (List Biologicals, 103B) with a 10-μL syringe (Hamilton) into the left sciatic nerve 4 days prior to euthanasia (day 24 after injury). On day 28, mice were perfused with 4% PFA and the spinal cords were postfixed for 1 week at 4°C and later cryoprotected in 30% sucrose. Serial tissue sagittal cryosections (50 μm thickness) were collected for free-floating immunohistochemistry. Sections were incubated overnight at 4°C with goat anti–CT-B primary antibody (1:30,000; List Biologicals, 703).

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

… 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