Bacterial Toxin Research Citations

Induction of EAE, evaluation of clinical disease, and administration of peptides:

Female SJL/J mice (6 to 8 weeks old) were purchased from Jackson Laboratories (Bar Harbor, ME) and housed in standard SPF facilities. On day 1, SJL/J mice were injected with 300 µg/mouse bovine myelin basic protein (Invitrogen) emulsified in Complete Freunds Adjuvant with 8 mg/ml H37Ra Mycobacterium tuberculosis (Sigma-Aldrich) and injected subcutaneously into two sites at the base of the tail along with 400 ng/mouse pertussis toxin (List Biological Laboratories Inc) in PBS i.p. On day 3, the pertussis toxin injection was repeated (Jager et al., 2011). …

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 evaluation of disease:

Experimental autoimmune encephalomyelitis was induced in C57BL/6 wild type mice by s.c. injection over four sites in the flank with 200 µg/mouse of myelin oligodendrocyte glycoprotein peptide 3555 (C S Bio Co. Menlo Park, CA, USA) in an emulsion with Complete Freunds Adjuvant (Difco, Detroit, MI, USA). Pertussis toxin (List Biological Laboratories Inc., Campbell, CA, USA) dissolved in PBS was injected i.p. at 200 ng/mouse at the time of immunization and 48 h 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)

EAE:

… C57BL/6 mice were obtained from the Centre for Animal Resources (CARE) of the National University of Singapore, and CD137L-/- mice were a gift from Amgen and bred in-house under pathogen-free conditions. Mice were injected subcutaneously with 100 µg MOG35-55 (Sigma-Aldrich) and 1 mg heat-killed Mycobacterium tuberculosis H37RA (Difco) emulsified in complete reunds adjuvant. Pertussis toxin (200 ng in PBS; List Biological Laboratories) was injected intraperitoneally on days 0 and 2 after immunization. EAE clinical symptoms were scored daily as follows: …

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)

… Cholera toxin (25 µg/mL stock; List Biological Laboratories, Campbell, CA). …

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

Liver Metastasis Model:

Tumor cells (5 10⁵/100 L phosphate-buffered saline [PBS]) were injected into the spleens of C57BL/6, CCR2 knockout (KO), severe combined immunodeficiency (SCID), or CD11b-diphtheria toxin receptor (DTR) mice. …CD11b-DTR or C57BL/6 mice received diphtheria toxin (7.5 ng DT/g body weight; List Biological Laboratories)…

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

Materials:

Recombinant light chain, purified 150 kDa BoNT/A, and SNAPtide (FITC/DABCYL) were from List Biological Laboratories (Campbell, CA), …

The above materials were used in the following:

• PL63 and PL50 Kinetic Parameters
• Fluorescent and HPLC Analysis of Peptide Cleavage and Impact of Exosite-Binding Peptides
• Standard BoNT/A LC Toxin Detection Curves
• Limit of Detection and Quantitation Determinations
• Effect of TMAO on Peptide Substrates

• Product #610A – Botulinum Neurotoxin Type A Light Chain, Recombinant
• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin

…After BDA administration, another glass capillary (tip diameter: 30 m) filled with 0.5% CTB (List Biological Laboratories, Campbell, CA) was lowered vertically into either the occipital cortex, area 2, lateral part (Oc2L), Te2D, or the temporal cortex, area 1 (Te1) (anteroposterior …

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

In vitro BoNT LC metalloprotease activity:

Determination of BoNT/A LC percent inhibition by SMNPIs was performed as previously described.  For BoNT/B LC percent inhibition, the HTS-assay utilizes a commercially available fluorogenic substrate from List Biological Laboratories (Campbell, CA). Briefly, a multichannel pipette was used to add fluorescent substrate (final concentration 20 M) and small molecules (final concentration 20 M) to a 96 well microplate. The reactions were initiated by adding recombinant BoNT/B LC (final concentration 40 nM) to each well. All reactions were conducted in a buffered solution consisting of 40 mM Hepes pH 7.2, 1 mM DTT and 100 M ZnCl2. The change in fluorescence intensity over time was continuously monitored in each well using a 96 well plate fluorimeter (Saphire 2, Tecan, Mnnedorf, Switzerland). The assays were run at 37 C, quenched by the addition of TFA, and analyzed using reverse-phase HPLC. Potential SMNPIs of the BoNT B LC were identified by comparing the reaction velocities (change in fluorescence intensity over time) of samples assayed in the presence of compounds versus  control samples.

Author did not specify which List Biological Laboratories product was utilized.  Refer to the follwing List Labs link for Botulinum products available:  https://listlabs.com/product-information/botulinum-toxins/

Cell treatments:

For Western blot experiments and luciferase assays the following treatment regimen was used. Six hours prior to harvesting, cells were treated with 10 mg/ml medium of Escherichia coliderived LPS (E. coli 055:B5; List Biological Laboratories, Campbell, CA), …

NOTE:  At the time this paper was written, Product #203 (5mg – LPS from Escherichia coli O55:B5) was utilized.

Product #203 (5 mg – LPS from Escherichia coli O55:B5) is no longer sold.

Product #203A (2.5 mg – LPS from Escherichia coli O55:B5) is available for purchase.

• Product #203A – LPS from Escherichia coli O55:B5

rPA Delivery in Rats:

Recombinant protective antigen from Bacillus anthracis (rPA) was obtained from List Biologicals (Campbell, CA). The MSA containing rPA was fabricated using a casting and drying fabrication method. …

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