Citations

Tracer injection:

Five days before being killed, the animals (rats) were injected into the sciatic nerve (11 bilaterally, 30 unilaterally) with a 1% wt per vol solution of the transganglionic tracer CTB (List Biological Laboratories, Campbell, CA) using a 36-ga NanoFil syringe. The sciatic nerve was exposed as described above and 1 µl was injected into each the tibial and common peroneal branches bilaterally (4 µl total).

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

EAE induction and antibody treatment:

Active EAE was induced by subcutaneous immunization of mice with an emulsion of 10 µg of MOG₃₅₅₅ peptide in CFA supplemented with 40mg of M. tuberculosis extract H37 Ra (Difco). In addition, the animals received 200ng of pertussis toxin (List Biological Laboratories) i.p. on day 0 and 2 after 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)

… in Complete Freund’s Adjuvant (Sigma-Aldrich). At the same time mice were also administered 250 ng of pertussis toxin (List Biological Laboratories) ip and, when indicated, again on d2. Clinical disease was monitored daily …

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)

BoNT-B:

The light chain of botulinum neurotoxin type B (50 kDa, List Biological Laboratories, Campbell, CA) was used to cleave recombinant Sb2.13

• Product #620A – Botulinum Neurotoxin Type B Light Chain, Recombinant

… CTB-labeled sensory axons can be identified by immunostaining with a goat anti-CTB IgG antibody (1:4,000, List Biological Laboratories, Campbell, CA, USA. # 703). …

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

Treatment of EAE by DiD-Labeled MSCs:

MOG₃₅₅₅ peptide (MEVGWYRSPFSRVVHLYRNGK, Peptron, Korea) was used to induce chronic EAE in C57BL/6 mice [18]. Briefly, 200 µg of MOG₃₅₅₅ diluted in 200 µL PBS and emulsified with 200 µL incomplete Freunds adjuvant (Gibco, Germany) and 1mg/mL heat inactivated mycobacterium tuberculosis (Difco, USA) in PBS was injected subcutaneously. After MOG₃₅₅₅ administration, 200ng pertussis toxin (List Biological Laboratory, USA) was injected intraperitoneally on the day of immunization and 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)

Induction and assessment of EAE:

For induction of EAE by active immunization, animals (mice) were injected i.p. with heat inactivated PTx (List Biological Laboratories) on days 0 and 2.

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)

Animals and vaccinations:

For rabbit experiments employing Freunds adjuvant, Female New Zealand white (NZW) rabbits (Covance Research Products, Denver, PA) weighing approximately 2.5 kg were immunized on day 0 with a mixture of two MAPs (125 g per MAP) in CFA and boosted 4 times at two-week intervals with the MAPs in IFA. For studies using human-use adjuvants, NZW rabbits were injected with TB-LND(2) or PA83 (List Labs, Campbell, CA) according to the above schedule, …

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

Reagents:

LPS was obtained from List Biological Laboratories (San Jose, CA, USA). …

EEM inhibits NF-B activation induced by LPS:

… Figure 1. EEM inhibits NF-B activation induced by LPS.  … (C) RAW264.7 cells were pretreated with 50 or 100 g/mL EEM for 1 h and then further stimulated with LPS (10 ng/mL) for 30 min. Cell lysates were analysed for IB and -actin protein by immunoblots. …

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

Injection of purified toxins:

HIOs were prepared and injected as described above. For these experiments, purified C. difficile toxin A (TcdA) or B (TcdB)…152C or 155D, respectively; List Biological Laboratories), reconstituted at a concentration of 1 g/ml, aliquoted, and stored at 80C. Due to lot-to-lot variation, each new lot was tested for toxin activity using the Vero cell cytotoxicity assay. Each HIO was injected with approximately 2 l of either 12.8 ng/l TcdA or 25.6 ng/l TcdB with 1 mM FD4. For each experiment, a new aliquot of toxin was thawed and used.

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