Citations

Surgery and tracer injections.

… Te posterior aspect of the skull was exposed by means of a skin incision and, depending on the experiment, a craniotomy was performed to expose either the cortex overlaying the SC or the PBG. A glass pipette (tip diameter 15–20 μm) containing CTB (List Biological Laboratories, Campbell, CA; 1% in phosphate bufer
0.1 M) was lowered vertically into the SGS layer of the SC or into the PBG, depending on the case. Collicular CTB injections were directed toward either the medial or the lateral aspect of the SC. …

Immunohistochemistry.

Immunohistochemical procedures were performed to reveal the expression of the choline acetyltransferase protein (ChAT) and the transport of the neural tracer cholera toxin subunit b (CTB) as reported in Deichler et al.42. All incubation steps were performed at room temperature in gentle agitation and preceded by PBS washes (3×5 min). First, to quench endogenous peroxidase activity, brain sections were incubated in 0.3% H2O2 and 10% methanol in PBS for 10 min. Next, the tissue was incubated overnight
at 4 °C with a primary antibody (1:20000 for anti-CTB [List biological Laboratories, RRID: AB_10013220] …

Author did not specify which CTB was utilized. List Labs has provided Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae) and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt). The former has been discontinued. The latter may be substituted for the former, if the researcher takes into account the difference in the buffer in which each product is provided and the amount of protein per vial. When Product #103B was reconstituted with 200 μl of water, it contained 1 mg of protein in 0.05 M Tris, 0.2 M NaCl, 0.001 M Na2EDTA, 0.003 M NaN3, pH 7.5. When Product #104 is reconstituted with 0.25 ml water, it contains 0.5 mg of Cholera Toxin B Subunit (choleragenoid) in 0.01 M Sodium Phosphate at pH 7.5.

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

… Bacillus anthracis protective antigen (List Biological Laboratories, cat. no. 171E) …

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

2.1. Induction of EAE with Myelin Oligodendrocyte Glycoprotein (MOG)35–55 Peptide

Reagents for EAE induction were prepared by diluting MOG35–55 peptide (2 mg/mL; Cat #: AS-60130-5; AnaSpec, Fremont, CA, USA) in UltraPure™ DNase/RNase-Free distilled water (ThermoFisher, Waltham, MA, USA) and emulsifying with equal parts of Complete Freund’s Adjuvant (BD Biosciences, Franklin Lakes, NJ, USA) containing 8 mg/mL Mycobacterium tuberculosis H37RA (Cat #: 231131; BD Biosciences, San Jose, CA, USA) by passage through glass Luer-Lok syringes and a micro-emulsifying needle for 45 min. The emulsion was then transferred to 1 mL Luer-Lok syringes with 27G ½” needles. Pertussis toxin was diluted in UltraPure™ water (2 ng/μL; Cat #: 181; List Biologicals Laboratories, Campbell, CA, USA) and transferred to syringes as described above. …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

… For pyrin inflam- masome activation, cells were resuspended in Opti-MEM (Life Technol- ogies) and stimulated with C. difficile supernatant (AB 2 [control] or AB+) for 12-16 h. Alternately, cells were stimulated with 0.2 mg/ml TcdB (List Biological Laboratories)…

• Product #155 – Toxin B from Clostridium difficile

Induction of EAE model

… Each rat was immunized by subcutaneous injection of antigen emulsion at a dose of 0.75ml/kg into its footpad, followed by an auxiliary injection of 10ug/kg diluted in 0.5ml pertussis toxin (List Biological Laboratories) in the dorsum of the foot. …

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)

… pathogenicity and toxin production. 0.5 ml commercially purchased purified CT (LIST Biological Laboratories, 101B, …

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

… To label abdominal premotor neurons in the caudal ventral respiratory group (cVRG), 6 rats were microinjected with the retrograde tracer cholera toxin subunit B (CTB, List Biologicals …

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

… Test sera were duplicated and diluted two-fold starting with a 1:50 PA 500 ng/mL in lethal factor (LF) 400 ng/mL (List Biological Laboratories Inc., USA). …

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

Induction of EAE and Clinical Assessment of Animals

Active EAE was induced according to a standard protocol [35]. Mice were immunized on day 0 by subcutaneous injection on both sides of the tail base with 100 μL of an emulsion composed of either PLP139-151 or MOG35-55 peptide in an equal volume of Freund’s adjuvant, supplemented with 4 mg/mL killed M. tuberculosis H37Ra (Difco Laboratories, Detroit, MI). The initial dose of antigen was 100 μg of PLP peptide for inducing relapsing EAE in SJL/J mice, or 200 μg of MOG peptide per GFAP-luc/B6(Cg)-Tyrc-2J/J mouse for induction of acute monophasic EAE. GFAP-luc/SJL/J mice received a second PLP antigen immunization on day 7. Immediately following these immunizations, all mice received intravenous (IV) injections of 200 ng Bordetella pertussis toxin diluted in 200-μL phosphate buffered saline (PBS) (List Biological Laboratories, Campbell, CA). The GFAP-luc/B6(Cg)-Tyrc-2J/J mice received an additional IV injection of 200 ng Bordetella pertussis toxin on postinoculation 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)

… according to the manufacturer’s instructions with 300 nM recombinant light chain from BoNT/C (#625A, List Biological Laboratories). After electroporation, the keratinocytes were washed and then seeded with sensory neurons …

• Product #625A – Botulinum Neurotoxin Type C Light Chain, Recombinant