Bacterial Toxin Research Citations

Pilocarpine and DT treatment:

At 8 weeks of age (1 week after the last tamoxifen injection), mice were injected intraperitoneally with 1 mg/kg methyl scopolamine nitrate… NestinCreER^T2::DTr^+/::GFP^+/ and NestinCreER^T2::DTr^/::GFP^+/ littermates that successfully entered and survived status epilepticus were selected to receive either DT (List Biological Laboratories, catalog #150, 40 g/kg, i.p.) or saline treatment. …

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

Chicken polyclonal IgY (#754A) against TcdB was purchased from List Biological Labs. …

Tracing:

Three days prior to terminal electrophysiology experiments, we retrogradely traced ulnar (flexor) and radial (extensor) motor neurons. Rats were anaesthetized and the axilla on the treated side was shaved and swabbed with alcohol. …2 µl of 1% Cholera Toxin beta subunit (CTb, List Biological Laboratories) in 0.9% saline and …

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

Lethal Toxin, consisting of protective antigen and lethal factor were from List Biological Laboratories. …

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

… 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