Citations

Materials and Methods:

The principal experiments were performed on 34 adult, male Lister Hooded rats weighing 270-320 g (Envigo). … Additionally, unconjugated CTB (List Biological Laboratories Inc., product #103B) was used as it is transported along axons in both anterograde and retrograde directions. This tracer was visualized by immunofluorescence. The tracer pairings were as follows: FB + FG, n = 6; CTB₄₈₈/CTB₅₉₄ + FB, n = 4; CTB in MBs + FB in RSP, n = 5; FB in MB + CTB in RSP, n = 2. Single tracer studies using only CTB were also conducted: CTB in RSP, n = 3; CTB in MB only, n = 4. …

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

EAE:

For chronic progressive EAE, C57BL/6 mice were immunized with 200 µg of recombinant myelin oligodendrocyte glycoprotein peptide MOG35 -55 … Intraperitoneal (i.p.) injections of 400 ng pertussis toxin (List Biological Laboratories) were performed on the day of immunization and on day 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)

After lidocaine HCL was injected into the scalp, an incision was made to retract the scalp and perform a craniotomy and duratomy on either the left or right hemisphere. Cholera toxin B subunit (CTb: List Biological Laboratories, Campbell, CA, …

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

EAE immunization:

… Female C57BL/6 mice (1012 weeks old) were immunized subcutaneously with 100 µg of myelin oligodendrocyte glycoprotein fragment₃₅₅₅ (MOG₃₅₅₅) peptide … Mice received an intravenous injection of 100 ng pertussis toxin (List Biological Laboratories Inc.) 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)

… Cholera toxin (Lot#10067A1) was obtained from List Biological Laboratories, Inc. (Campbell, CA, USA) 

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

Materials:

… lyophilized LPS from E. coli J5 strain was purchased from both List Biological Laboratories, Inc …

• Product #301 – LPS from Escherichia coli J5 (Rc)

Materials and Methods

Anthrax lethal factor (Product #172), and the chicken IgY polyclonal anti-LF antibody (Product # 769A) are
products of List Biological Laboratories, Inc. The Nunc-Immuno Tubes, Maxisorp (cat# 444202) used for LF
antibody coating and the dimethyl sulfoxide (DMSO) (cat # TS-20684) were purchased from ThermoScientific.
The 96-well, black, flat bottom, non binding plates used for the fluorescent plate assay were from Corning
(cat # 3991). Bovine plasma (cat # 7310806) was purchased from Lampire Biological Laboratories.

Sample Preparation: Stock solutions of the fluorogenic substrates, MAPKKide® Plus (± biotin), were made
2.5 mM in DMSO based on the peptide content determined by elemental analysis. The substrates were diluted
in assay buffer: 20 mM HEPES, pH 8.0 containing 0.1% Tween-20. The LF was added to neat bovine plasma.

LF Activity Assays:
Antibody Capture: The Nunc-Immuno Maxisorp Tubes were coated with 1 mL of a 10 μg/ml solution of
a chicken affinity purified polyclonal IgY antibody to anthrax lethal factor (List Prod # 769A).
The immuno-tubes were incubated with the IgY overnight at 2-8°C. After 5 washes with 1.5 mL each of PBS containing 0.05%
TWEEN-20 (PBST), the anti-LF coated tubes were exposed to 1 mL of a series of LF concentrations in neat
plasma. The tubes were incubated at 22°C for 1 hour. Tubes were then washed 3 times with 1.5 ml PBST and
1.0 ml of 10 μM MAPKKide® Plus (± biotin) was added. The reaction was allowed to proceed for 3, 5 and
22 hours at 37°C. At each time point 250 μl of the reaction mixture was removed from triplicate tubes and placed
in a 96 -well black plate. In the case of the biotinylated MAPKKide® Plus a 0.5 ml aliquot of each triplicate
was added to streptavidin-coated beads after 22 hours of exposure to LF. The mixture was gently rotated at
ambient room temperature for 1 hour and then read in the platereader.

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #769B – Anti-Lethal Factor from B. anthracis (Chicken lgY)
• Product #532 – MAPKKide® Plus (AMC) Specific Substrate for Anthrax Lethal Factor

… All drugs for mucosal immunization were diluted in 300 µL PBS. Two oral immunizations with 10 µg CT (List Biological Laboratories, Campbell, CA) were administered on day 0 and 10. …

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

… Mice also received 300 ng of Pertussis toxin (List Biological Laboratories) iv on the day of immunization and 2 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)

Animals and EAE mice induction:

C57BL/6 mice, 68-week old, female, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. IL10^/ mice, 68-week old, female, were purchased from Model Animal Research Center of Nanjing University. Those mice were immunized subcutaneously with 400 µg MOG₃₅₅₅ peptide (Sangom Biotech) emulsified in CFA (Sigma) containing 4mg/ml heat-killed Mycobacterium tuberculosis H37Ra (BD Biosciences) on day 0. In addition, 200ng pertussis toxin (List Biological Laboratories) in 0.1ml PBS was administered intravenously on day 0 and day 1 post immunization per mice. …

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)