Bacterial Toxin Research Citations

Misoprostol and LPS administration

For each treatment arm, manure was manually evacuated prior to administration of misoprostol (M-PO, M-PR) or water (CON). For per os administration (M-PO), misoprostol tablets (100 μg; Lupin Pharmaceuticals) were dissolved in 30 mL of water and administered via oral syringe. This was followed by administration of 30 mL of water through the same syringe for a total volume of 60 mL delivered. Water (60 mL) was also administered per rectum via 16-inch 8-French red-rubber catheter advanced approximately 30 cm into the rectum. For per rectum administration (M-PR), misoprostol tablets dissolved in 30 mL of water were infused via red-rubber catheter as described above, followed by infusion of 30 mL of water through the same syringe for a total volume of 60 mL delivered. Water (60 mL) was then administered orally via syringe. Horses in the CON treatment received water per os (60 mL) and per rectum (60 mL) as already described. Immediately following treatment administration (M-PR/M-PO/CON), LPS (10 µg/mL; List Biological Labs) was administered IV at 30 ng/kg diluted in 500 mL saline (0.9% NaCl) solution as a 30-minute continuous rate infusion.

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/

Optic nerve crush and intraocular injections

… All viruses were injected in a volume of 3 µl and a titer of 1 × 1013 gc/ml 2 weeks prior to optic nerve crush to insure adequate time for gene deletion or transgene expression at the time of nerve damage. Two days prior to the end of a 14-day survival period, cholera toxin B subunit (CTB, 3 µl/eye, 2 µg/µl, List Biological Laboratories, Inc., 103B) was injected intraocularly as an anterograde tracer to label axons regenerating through the optic nerve. …

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

Maintenance culture conditions

All MCF10A-derived cell lines were maintained in standard MCF10A growth medium without antibiotics (DMEM/F12 medium (Wisent Bioproducts, 319-075-CL), 5% horse serum (Gemini Bio, 100508), 10 µg ml−1 insulin (Thermo Fisher Scientific/Gibco, A11382II), 0.5 mg ml−1 hydrocortisone (Sigma-Aldrich, H4001), 20 ng ml−1 EGF (R&D Systems, 236-EG-01M) and 100 ng ml−1 cholera toxin (List Biological Laboratories, 100B)). …

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

EAE and EAE tissue isolation

Female C57BL/6 mice (8–10 weeks old) from Jackson Laboratories were immunized subcutaneously with MOG 35–55 peptide (50 μg/100 μl, Protein and Nucleic acid facility, Stanford University School of Medicine, Stanford, CA) emulsified in complete Freund’s adjuvant (CFA) (Thermo Fisher Scientific) containing 10 mg/ml of heat inactivated Mycobacterium tuberculosis H37RA (Sigma-Aldrich). 50 μl emulsion was injected at one site into each hind flank. On the day of immunization and 48 h later, animals received intraperitoneal injections of 300 ng of pertussis toxin (List Biological Laboratories). …

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)

… The assay mixtures were allowed to incubate at rt for 30 min, before the addition of 25 µL of SNAPtide substrate flP6 (List Laboratories) to initiate the enzyme reaction, …

• Product #523 – SNAPtide® Peptide Substrate flP6(DABCYL/5-IAF) for C. botulinum Type A Neurotoxin

2.1 Materials and Chemicals

… The aLF was ordered from List Biological Laboratories, Inc (Listlabs, US). …

2.2 The Preparation of aLF Proteolytic Reaction

aLF, ZnCl2 and peptide substrate were mixed together as proteolytic buffer (5.64 μM, 28 mM and 2.82 μM) for endopeptidase activity reaction, …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Purified LPS and membrane receptor blots

LPS was purified by the hot phenol method previously described41. Purified LPS from E. coli 0111:B4 and Salmonella typhimurium were purchased from List Labs. …

• Product #201 – LPS from Escherichia coli O111:B4
• Product #225 – LPS from Salmonella typhimurium

Induction of active EAE

SJL/J 68-wk-old females (The Jackson Laboratory, Bar Harbor, ME) were actively immunized with PLP 139151, but otherwise
maintained, handled, and surveilled as outlined previously (25). Briefly, mice were actively immunized by s.c. injection of 0.2 ml inoculum containing 1030 mg PLP 139151 (HSLGKWLGHPDKF) peptide in IFA (Difco Laboratories, Detroit, MI) with 800 mg
Mycobacterium tuberculosis hominis H37Ra on days 0 and 7 (26). These inoculations were followed with 200 ng pertussis toxin i.p. (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)

Immunolabeling

The following primary antibodies were used: … CTB 1:5000 (List Biologics 703 lot# 7032A10) …

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

… ELISpot plates (Millipore) were coated with PT (0.1 µg/µl, List Biological Technology) and anti-human IgG (15 µg/ml, Biolegend) overnight at 4 °C. Coated plates were washed and blocked with RPMI medium 1640 basic …

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)