Citations

Chemicals and reagents:

All bacterial toxins were purchased from List Biological Laboratories (Campbell, CA). …

Human B-lymphocytes sensitivity to Pseudomonas toxin-mediated lethality:

Human B lymphocytes were treated with serial dilutions of P. aeruginosa exotoxin A for 48hours.

Cellular drug screens:

RAW264.7 cells (10,000 per well) were seeded in 96-well plates 24hours before the assay. Cells were treated with compounds for 1hour, and then challenged with either 2g/ml PE or 4g/ml cholera toxin for 12hours. As rodent cells are insensitive to diphtheria toxin, C32 cells were treated with 2g/ml of diphtheria toxins for 24hours. …

Toxins treatments and cell viability assays:

Cells (10,000 per well) were seeded in 96-well plates 24hours before the assay. Cells were treated with Bithionol for 1hour. RAW264.7 cells were challenged with anthrax toxins that include LF or FP59 and PA83 or PA63 (for 6hours), PE (for 12hours), or cholera toxin (for 12hours) at 0.5, 2, and 4g/ml respectively. C32 cells were treated with 2g/ml of diphtheria toxin for 24hours. Determination of cells viability was performed by MTT assay….

MAPKK2 cleavage assay:

… Following pre-treatment, the cells were exposed to 0.5g/ml of PA and LF at 37C for up to three hours in the presence of 33M of Bithionol. …

Cellular cathepsin B and caspase-1 activity assays:

…Caspase-1 activities were induced by 1hour pretreatment of cells with 0.5g/ml of LF+PA, and was compared to cells un-induced by the toxin. …

BoNT/A light chain FRET assay:

The reaction volume was 250l per well in 96 wells plate, containing 50mM HEPES pH 7.4 containing 0.05% TWEEN 20, 5M SNAPtide (BoNT/A substrate peptide) conjugated with DABCYL and FITC (List Biological Laboratories, Inc), and 33M of Bithionol. …

• Product #160 – Exotoxin A from Pseudomonas aeruginosa
• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).
• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis
• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin
• Author did not specify which Cholera toxin was utilized.  List Labs provides Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae and Product #101 – Cholera Toxin from Vibrio cholerae.

Reagents and antibodies:

…Cholera toxin B subunit (CTxB) biotin conjugate was from List Biological Laboratories …

FACS analysis:

Cell surface expression of gangliosides was analysed by flow cytometry. The following antibodies/agents were used to detect cell surface gangliosides; GM3, M2590; GM2, 1011; GM1, biotin-conjugated CTxB, …

• Product #112 – Cholera Toxin B Subunit Biotin Conjugate

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

EAE disease model:

On day 0, 8-week-old male mice were immunized subcutaneously with 50µg MOG₃₅₅₅ peptide emulsified in complete Freunds adjuvant …On days 0 and 2, 150ng per mouse pertussis toxin (List Biological Laboratories) was given by intraperitoneal injection. …

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)

EAE induction and clinical score:

EAE was induced in female C57BL/6 mice…200 ng pertussis toxin (List Biological Laboratories Inc.) was administered intraperitoneally on day 0 and 2. MOG_35-55 induced EAE mice were treated daily with 10 mg/kg clemastine (in PBS with 10% DMSO by oral gavage) or the equivalent volume of vehicle. …

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)

Clostridium difficile exotoxin TcdA List Laboratories 152C …

• Product #152C – Toxin A from Clostridium difficile

Immunization:

DT or its non-toxic mutant CRM197 (List Biological Laboratories, Campbell, CA) was injected i.p. (0.5 g in 0.5ml PBS per dose, except for CD11c-DTR;Mgl2-DTR mice in which only a single dose of 125 ng per mouse was given to avoid toxicity) at indicated time-points. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #149A – Diphtheria Toxin CRM₁₉₇, Mutant

… Six-week-old female specific-pathogen-free BALB/c mice, purchased from Charles River Laboratories (Saint Germain sur l’Arbresle, France), were sensitized with 1 mg of either OVA, OM or LYS, plus 10 µg of cholera toxin (CT, List Biologicals, Campbell, USA),…

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

Reaktivita imunitnho systmu byla podpoena intraperitoneln aplikac 0,6 a 1,2 g PT (pertusov toxin; List Biologicals, Campbell, USA), rozputnho v PBS (phosphate buffered saline).

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)

…Recombinant protective antigen (rPA) was purchased from List Laboratories. …

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