Citations

EAE induction

EAE was induced in mice by subcutaneous injection of 200 μg MOG35–55 peptide with 500 μg M. tuberculosis strain H37Ra (Difco) emulsified in incomplete Freund Adjuvant oil in 200ul volume into the flanks at two different sites. In addition, the mice received 400 ng pertussis toxin (PTX; List Biological Laboratories) i.p. at the time of immunization and 48 h 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)

Induced experimental autoimmune encephalomyelitis (EAE) model

Per mouse, 200ug of MOG20,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54 peptide (EZ Biolabs #cp7203) emulsified in Freund’s Complete Adjuvant (Santa Cruz Biotechnologies #sc-24018). Mice were then subcutaneously injected with 150ul of the emulsification with 75ul each on the right and left flanks. Mice were then intraperitoneally injected with 200ng of pertussis toxin (List Biologicals #180) and then once again two days later. Mice were assessed daily for clinical signs of EAE. …

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

… About 500 ng of pertussis toxin (List Biological Laboratories, Campbell, CA) was injected intraperitoneally on day 0 and 2. Mice were monitored for body weight and EAE onset 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)

… Pertussis toxin (100 ng/mouse, List Biological Laboratories) was administered intravenously into the tail vein on day 0 and day 2 after immunization. The disease course was monitored daily, and mice were assigned a clinical score as …

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)

… goat anti-EF (773L, List Labs, Campbell, CA, USA), …

… Pertussis toxin (200 ng; List Biological Laboratories, Inc., Campbell, CA, USA) was intravenously administered at 0- and 48-h post immunization. EAE scores were assessed daily according to the …

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 addition, mice were intraperitoneally administered 500 ng pertussis toxin (PTX, List Biological Laboratories, California, USA) on the day of immunization and …

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)

2.3. Study design and active immunization

Action immunization was conducted as previously reported [16]. The peptides were dissolved in saline (2 mg/ml), and then fully
emulsified in an equal volume of complete Freund’s adjuvant containing 1 mg/ml heat-killed Mycobacterium tuberculosis H37Ra
[(CFA), #F5881, Sigma Aldrich, USA]. Mice were randomly assigned to experimental, negative control, or positive control groups.
Each mouse received a subcutaneous injection of emulsion (200 μl containing 200 μg of peptide) at four sites on the back, which was
repeated three times at 2, 4, and 6 weeks (Fig. 1). The GluN1359-378 peptide emulsion was injected as the positive control. For the sham
groups, an equal volume of saline emulsified in CFA was used in place of the peptide. Two hundred nanograms of pertussis toxin (1 ng/μl, #181, List Biological Laboratories, USA) was intraperitoneally administered to each mouse at the time of the last immunization and 48 h 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)

Silver staining and western blot analysis

TcdA (1 μg) (List Biological Laboratories, Inc. California, USA) was incubated with DMC-4 (35 μl) for 1 h at 37 °C. Samples were loaded using 6X SDS loading buffer and heated at 95 °C for 10 min. All samples were loaded on to gradient SDS-PAGE gels (4-15%) after boiling. Silver staining was performed using the Pierce Silver Stain Kit as per the manufacturer’s instructions (Thermo Fisher Scientific. USA).

TcdA and TcdB (100 ng) were incubated with DMC-4 (35 μL) for 1 h at 37 °C. Samples were loaded using 6X SDS loading buffer and heated at 95 °C for 10 min. All samples were loaded into gradient SDS-PAGE gels (4-15%) after boiling. …

• Product #152C – Toxin A from Clostridium difficile

… Three weeks after the completion of tamoxifen treatment, mice were given two intraperitoneal (i.p.) injections (three days apart) of 40 µg/kg of diphtheria toxin (DTX, List labs, 150) in saline. Three weeks after DTX treatment, mice were inoculated with IAV as described above …

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