Bacterial Toxin Research Citations

Bordetella pertussis toxin (salt-free) List Biological Laboratories, Inc. Cat # 181

Active EAE
Eight to 12-week-old female and male Cst3−/− and C57BL/6J WT littermates were immunized subcutaneously with 100μg myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) (Stanford Pan Facility) mixed in a 1:1 emulsion with complete Freund’s adjuvant that contained 4mg/ml Mycobacterium Tuberculosis H37Ra (Fisher Scientific). Animals were also administered 400ng of Bordetella Pertussis toxin (PTX) (List Biological Laboratories Inc.) intravenously on days 0 and 2 post-immunization. …

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

EAE

Mice were immunized subcutaneously in four sites on the back with 100 µg MOG(35–55) peptide (Biosynthesis) emulsified with CFA with M. tuberculosis strain H37Ra (DIFCO Laboratories). Mice also received 100 ng pertussis toxin (List Biological Laboratories) intraperitoneally on days 0 and 2. Mice were assessed daily and scored as follows: 1, flaccid tail; 2, impaired righting reflex and hindlimb weakness; 3, partial hindlimb paralysis; 4, complete hindlimb paralysis; 5, hindlimb paralysis with partial forelimb paralysis; or 6, moribund.

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)

… Control containing purified C. difficile toxins (A and B; List Biological Labs, CA, USA) and antitoxin were included. Plates were viewed under 200 magnification for Vero cell rounding after an overnight incubation at 37°C/5% CO2 …

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

Diphtheria Toxin Biological List Laboratories Cat #150

… To induce Cre positive cell deletion, Diphtheria toxin (Biological List laboratories) (100ng/mouse/day) was daily i.p. injected into mice. Cre negative littermate got same amount of DT as control. …

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

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