Bacterial Toxin Research Citations

2.6. Toxin Neutralisation Titre (TNA) for Neutralising Antibodies Titre

The in vitro toxin neutralisation ability of vaccinated cattle antibodies was determined as previously described by Ndumnego et al. [38]. Briefly, the TNA was determined with 1.0 × 105 of J774A.1 mouse macrophage cells (ECACC cat no 91051511) per well followed by overnight incubation at 37 °C and 5% CO2. Then sera from vaccinated animals were 2-fold serially diluted in duplicates starting from the dilution of 1:50. Then the macrophages were exposed to anthrax lethal toxin (PA: 500 ng/mL and LF: 400 ng/mL (List Biological Laboratories Inc., Campbell, CA, USA) in the presence of antibodies. …

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

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

Intersectional genetic ablation of Tac2 neurons in the spinal cord

Triple mouse line (Tac2Cre, Lbx1flpo and TauDTR mice) was generated for cell ablation. To ablate Tac2 neurons in the spinal cord, i.p. injection of diphtheria toxin (DTX, 40 μg kg−1, List Biological Lab) was injected at day 1 and day 4, respectively41. Behavioral and histochemical experiments were performed 2 weeks after the second DTX injection.

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

hMSC treatment with cytokines or LPS

For these experiments, hMSCs were plated at 1.6 x 105 cells/cm2 on 60mm untreated tissue culture plates in DMEM-F12 supplemented with antibiotics. TNF-α, IL-1β, and IFN-γ were obtained from R&D Systems. As previously described, CytoMix consisted of 50 ng/mL TNF-α, IFN-γ, and IL-1β [12]. For cytokine array experiments, TNF-α, IFN-γ, and IL-1β were used at 50 ng/mL in various combinations. LPS concentrations ranged from 5 pg/mL– 1 mg/mL (List Labs or Sigma). In all cases, the hMSCs were exposed to the experimental condition for 24 hours prior to harvesting, unless otherwise noted.

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/

Anatomical Tracer Injections
… During a sterile surgery, Lidocaine HCl was injected into the scalp prior to incisions. The scalp was retracted, and a craniotomy and durotomy were performed on either the left or right hemisphere. Cholera toxin B subunit (CTb: List Biological Laboratories, Campbell, CA, United States, Cat. no. 104) was reconstituted in 0.1 M potassium phosphate buffer (PB, 1%, pH 6.0) and either pressure-injected or delivered with current into area 18 (approximately 2.3 mm from the occipital pole). …

TABLE 1
Antibodies used in this study.

Antibody Source Manufacturer Dilution
Anti-Cholera Toxin B Subunit Goat polyclonal List Biological Laboratories, Campbell, CA, United States (Cat# 703) 1:5,000

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

EAE induction

C57BL/6 mice were injected subcutaneously in the flanks with 200 μL of 1 mM myelin oligodendrocyte glycoprotein (MOG)35–55 peptide (Peptide Synthesis Core Facility, University of Utah) emulsified with reconstituted complete Freund’s adjuvant (CFA), composed of incomplete Freund’s adjuvant (Pierce Biotechnology, Rockford, IL, USA) containing Mycobacterium tuberculosis H37 Ra (2 mg/mL) (Difco Laboratories, Detroit, MI, USA). Mice were injected intravenously with 100 μL Bordetella pertussis (BP) toxin (List Biological Laboratories, Inc., Campbell, CA, USA) at 0.2 μg per mouse on days 0 and 2 following sensitization. MOG35–55-induced EAE mice developed a monophasic disease course. …

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)

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)