Bacterial Toxin Research Citations

Experimental Materials:

Stx2 toxin was purchased from List Biological Laboratories, Inc. (Lot #1621A1, Campbell, CA). Endotoxin levels were tested by List Biological Laboratories and found to be acceptable. …

Determination of Mean Lethal Dose:

Groups of at least 10 randomly selected mice were treated by intraperitoneal (ip) injection with 500 L per dose of serial dilutions of Stx2 (in a range that spans high lethality to no deaths). …

Toxicokinetics of Stx2:

The biologic half-lives of Stx2 were determined in the presence or absence of mAbs against Stx2. Mice were treated iv with 100 ng per mouse (100 L of 1,000 ng/mL stock) of Stx2. …

Treatment of Mice Post-intoxication or Pre-intoxication with Stx2 mAbs:

To simulate post-intoxication treatment, mice were treated by iv with 100 L of 180 ng/mL of Stx2. …

List Labs has discontinued Product #162 (Lyophilized) and has replaced Product #162 with Product #162L (Liquid):

• Product #162L (Liquid) – Shiga Toxin 2 from Escherichia coli

…Lipopolysaccharide (LPS E. coli serotype 011:B4) was purchased from List Biological Laboratories (Campbell, CA).  …

Determination of TLR4 Ligand Presence in BAL Fluid and Serum:

…All conditions were tested in duplicate in the presence or absence of polymixin B (50 g/ml), which binds to LPS and prevents recognition by TLR4. Positive control wells were treated with LPS at 10 ng/ml with or without polymixin B. Negative control wells were treated with medium only.

• Product #201 – LPS from Escherichia coli O111:B4

Materials and Methods – Stx and monoclonal antibodies (mAbs):

Pure Stx1 was purchased from List Biological Laboratories, Inc. (Campbell, CA). …

Results – Development and characterization of a polyclonal antibody against Stx2:

…The pAb obtained bound to both the recombinant Stx2a toxoid and the wild type Stx2a but did not react with Stx1 when tested by direct ELISA (Table 3), indicating the pAb is Stx2-specific. …

• Product #161 – Shiga Toxin 1 from Escherichia coli

… Anthrax protective antigen-83 (PA83) was purchased from List Biological Laboratories. …

MPII Proteolysis of Proteins in Vitro:

Anthrax PA83 (2 g; 1 m) was co-incubated for 1 h at 37 C with increasing concentrations of MPII in 50 mm HEPES, pH 8.0, containing 1 mm CaCl₂, 0.5 mm MgCl₂, and 10 m ZnCl₂. The total volume of the reactions was 20 l. Where indicated, GM6001 (1 m) was added to the reactions to inhibit MPII. The cleavage reaction was stopped by adding a 5 SDS sample buffer. The digest samples were analyzed by SDS-PAGE using 420% polyacrylamide gel.

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

… SNAPtide (#521, List Biological Laboratories, Campbell, CA, USA) solution (60 L, 4.9 M) was incubated in the dark with LC/A1 in LC/A Hydrolysis Buffer (50 mM HEPES, 1 mg mL 1 BSA or 0.05% TWEEN 20, pH 7.4, titrated with HEPES potassium salt) at RT for 20 or 60 …

• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin

3D-EM of the L-PTC and the HA complex

The L-PTC of BoNT/A was obtained from List Biological Laboratories, Inc. (Campbell, California) and Miprolab GmbH (Gttingen, Germany). …For the L-PTC, no symmetry was imposed throughout the 3D reconstruction and refinement, while for the HA complex, a C3 symmetry was imposed. Refinement was terminated when no significant changes could be visually detected. A data set of 15,140 particles was used for the final reconstructed map of the L-PTC, for which the resolution was estimated to be 30.8 based on the resolution criteria of Fourier shell correlation (FSC) at 0.5 cutoff. …

In vivo immunization.

TNF/Rorc(γt)+/+ and TNF/Rorc(γt)−/− mice at 6–8 weeks were immunized with OVA (grade V; Sigma-Aldrich, St Louis, MO) by intragastric gavage of 100 μg of OVA+20 μg cholera toxin (List Biological Laboratories, Campbell, CA) on seven occasions at 7-day intervals. …

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

Chemicals and Reagents:

PA and LF were purchased from List Biological Laboratories…Batches of PA and LF had different potency, accounting for the different amounts used in different experiments, each of which included control data for the same toxin batch.

Toxin Treatment and Cell-Viability Assay:

Cells were seeded in a 96-well plate at a density of 2 10⁵ cells/mL 1 d before toxin treatment. Various concentrations of PA combined with a fixed concentration of LF (200 ng/mL) were added to the wells, and cells were incubated for 4 h at 37 C.

Immunofluorescence Microscopy.

PA protein was labeled with Alexa-Fluor 488 using the protein-labeling kit…

Biochemical Assay of PA Binding and Internalization.

Cells were exposed to 1 g/mL of PA at 4 C for 1 h for the binding assay and then were shifted to 37 C for the indicated time for the internalization kinetics assay.

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

Cell culture and differentiation:

CD4+ T cells were cultured for 24 d in serum-free X-VIVO 15 medium.  …CT (List Biological Laboratories), …were added at various concentrations to the culture media as indicated.

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

Tissue processing and anatomical reconstruction:

Spinal cord:

Before the spinal cord was removed, the level of the spinal cord lesion was identified and marked with pins contralateral to the lesion, one just rostral and the other just caudal. This enabled us to reliably identify the lesion in the processed sections, and also served as alignment landmarks during reconstruction. The perilesioned spinal cord was sectioned horizontally. An alternating series of horizontal sections was reacted for cytochrome oxidase, enabling clear demarcation of the gray and white matter. Another series was incubated with primary goat anti-CTB (1:4000; List Biological Laboratories, Campbell, CA) diluted in …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)