Bacterial Toxin Research Citations

… ELISA to assess cholera-specific IgG concentrations was performed by coating the plates with a final concentration of 1 µg/ml with the Vibrio (V.) cholerae strain Inaba 569B (List Biological Laboratories) and incubated for 3 h at 37°C. …

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

EAE:

…For EAE, 100 ng (or 200 ng for Fcr1 / mice) of pertussis toxin (List Biological Laboratories) was given ip on days 0 and 2 after immunization. Mice were monitored daily …

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 induction with MOG₃₅₅₅:

…EAE was induced by subcutaneously immunizing mice in the flanks with myelin oligodendrocyte glycoprotein peptide (MOG)_3555…Intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories) followed on the day of immunization as well as 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)

…Rats (6-7 weeks old, 260-300 g, Charles River, lArbresle, France) were anesthetized as described above and placed in a stereotaxic apparatus. Glass micropipettes (4-5m O.D.) were backfilled with a 1% solution of cholera toxin B subunit (CTb; List Biological Lab., diluted in 0.1 M phosphate buffer (PB) at pH 6.0) for injection in the dentate gyrus (n=4), the retrosplenial cortex (n=4) and the supramammillary nucleus (n=4). …

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

Cell stimulation:

Monocytes derived from patients with RA were plated in 96-well culture plates at 5.10⁴ cells/well. NI-0101, formerly designated as Hu 15C1 [17], or a human IgG₁ isotype control (clone DA4) was added to wells at a final concentration of 20 g/mL. Plates were incubated (30 minutes, 37 C) prior to adding SF (pooled or individual) diluted to a final concentration of 2.5 %. Ultrapure lipopolysaccharide (LPS) (Salmonella minnesota), added at 10 ng/mL (List Biological Laboratories), was used as a positive control. …

• Product #434 – ULTRA PURE LPS from Salmonella minnesota R595 (Re)

Chemicals and Reagents:

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

LF FRET Drug Screen and Data Analysis:

For screening JHCCL drugs in 96-well plates, the reaction volume was 250l per well, containing 20mM HEPES pH 7.2, 5M MAPKKide conjugated with DABCYL and FITC (List Biological Laboratories, Inc), and 33M of JHCCL compound. Reactions were initiated by adding anthrax LF…

Cell Culture, Toxins Treatments, and Cell Viability Assays:

RAW264.7 mouse macrophage cells were maintained in DMEM (Invitrogen) supplemented with 10% FBS (Corning) and 100g/ml penicillin and streptomycin. …Cells were treated with drug hits for 1hour at 37C 5% CO₂. RAW264.7 cells were challenged with anthrax toxins that include LF and native 83kDa PA (for 3hours), LF and PA in the 63kDa form (for 6hours), P. aeruginosa exotoxin A (for 12hours), or Cholera toxin (for 12hours), which were also pre-treated with identical drug concentrations, such that the final toxins concentrations were 0.5, 0.5, 2.0, and 4.0g/ml respectively. …

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

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis
• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Toxin Neutralization Assay (TNA):

… Goat sera (starting dilution; 1:50) were doubly diluted in culture medium containing PA and LF (List Biological Laboratories, Inc …

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

EAE:

…Pertussis toxin (PT, List Biological Laboratories, CA) was injected intraperitoneally on post-immunization days 0 and 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)

Retrograde tract tracing experiments:

The retrograde tracer, cholera toxin subunit (CTB, List Biological Laboratories, Campbell, CA), was injected by iontophoresis into the PB in three animals. …

Immunohistochemical identification of the CTB injection sites:

CTB injection sites were identified in single immunolabeled sections. One of the four series of sections was pre-treated sequentially, first with 0.5% H₂O₂ and 0.5% Triton X-100 in PBS for 15 min, and then 2% normal horse serum in PBS for 20 min. Sections were then incubated in a mixture of goat anti-CTB serum (List Biological Labs) at 1:10,000 dilution for 2 days at 4°C. …

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

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.