Bacterial Toxin Research Citations

Drug Delivery:

All drugs were made in sterile double distilled water (ddH₂0) containing 1% DMSO (100x) unless otherwise noted. Vehicle conditions are cells that were supplied with ddH₂0 and DMSO to a final amount of 0.01% per 3ml of media. After 24 h of transfection, media was removed from cells and replaced with the indicated drug for the length of 48 h unless otherwise noted. In the case of time-course experiments, agonists, modulators and/or secondary drugs (botulinum toxin b) were added for varying durations such that all time-based groups were harvested at the end of the 48 h drug incubation period. This procedure ensured that all groups had equivalent time for protein synthesis and that tissue for immumocytochemistry could be processed in parallel. Final concentrations were as follows: GABA (1 and 10 M), gaboxadol (10 M), -alanine (10 M, 50 M and 1 mM), THP (100 nM) and botulimum toxin B (5 nM). All drugs except for THP (Steraloids, Newport, RI) and botulinum toxin b (List Biological Laboratories, Inc., Campbell, CA) …

… Pressure injections of volumes ranging from 20 to 200 nl of CT (1% low salt solution, List Biologicals, Campbell, CA) were made with a syringe (external diameter 0.24 mm, Hamilton Co., Reno, NV) at a rate of 10 nanoliters per minute (Trojanowski, 1983, Ericson and Blomqvist, 1988). …

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

Materials and methods – Chemicals:

… Cholera toxin was purchased from List Biological Laboratories, Inc. (Campbell, CA, USA). …

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

Experimental autoimmune encephalitis (EAE) induction and clinical scoring:

Six to eight-week-old female mice were immunized subcutaneously with PLP₁₈₀₁₉₉ peptide (200µg/mouse, Genemed Synthesis Inc.) emulsified in CFA containing Mycobacterium tuberculosis H37Ra (500µg/mouse, BD Diagnostics). Mice also received 300ng of pertussis toxin (List Biological Laboratories) intraperitoneally (i.p.) at the time of immunization and 48 hours 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)

Synthesis of HD5 and Mutant Analogues:

… Recombinant anthrax lethal factor was purchased from List Biological Laboratories, Inc. …

LF Inhibition Kinetics:

The inhibition of LF by various defensins was quantified using an enzymatic kinetic assay (52). Briefly, freshly prepared LF at a final concentration of 1 g/ml (10 nm) was incubated at 37 C for 30 min with a 2-fold dilution series of defensin in 20 mm HEPES buffer containing 1 mm CaCl2 and 0.5% Nonidet P-40, pH 7.2. 20 l of LF substrate (1 mm) was added to each well to a final concentration of 100 m in a total volume of 200 l. The enzyme activity, characterized as a time-dependent absorbance increase at 405 nm due to the release of p-nitroaniline, was monitored at 37 C over a period of 5 min on a 96-well Vmax microplate reader …

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).

Antigen presenting cell cultures:

Adherent splenocytes, and resident peritoneal cells were cultured for 4 or 24 hours in the presence or absence of 100 ng/ml LPS (E. coli O111:B4; List Biological Laboratories, concentration previously determined).  After incubation, supernatants from each culture were collected for analysis by multiplex bead array, and the cells were harvested for analysis by flow cytometry.  

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

Mice and Immunizations:

… In CD11c-DTR mice, DCs were depleted by i.p. injection of 8 ng diphtheria toxin (DT) (List Biological Labs) per gram body weight, in combination with rear footpad injection of 10 ng of DT each, all 12 hours before VRP injection.

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

Phage display and phage ELISA:

… To measure the activity of phage-displayed scFvs, phage ELISAs were performed in high-binding 96-well plates (Costar) coated with 1 g/ml KJ126 monoclonal antibody (BD Pharmingen; to assess DO11.10 scFv activity), 16 g/ml anthrax protective antigen (List Labs; 14B7), …

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

Materials:

… Protective antigen was acquired from List Biologicals (Campbell, CA), …

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

Tissue processing and data analysis:

After 57 days of survival, galagos were sacrificed using an overdose of sodium pentobarbital (80 mg/kg) and perfused with phosphate-buffered saline (PBS: pH 7.4) followed by 2% paraformaldehyde, and finally with a solution of 2% paraformaldehyde with 10% sucrose. The brains were removed, the cortex was separated from underlying brain structures, the sulci were opened, and the brain was flattened as described … The cortical sections were divided into four series and processed for cytochrome oxidase (CO; Wong-Wiley, 1979), myelin (Gallyas, 1979), or CTB using a protocol described in Baldwin et al. (2011) (Table 1). The last series was not processed, but instead mounted directly onto slides and coverslipped for fluorescent analysis of neurons labeled with FR. The brainstem was processed in five series, which were CO, CTB, FR, and acetylcholinesterase (AChE: Geneser-Jensen and Blackstand, 1971); the fifth series was set aside and processed as part of another study. Here we describe only the results from cortex and the locations of injections in the SC.

… Table 1. Antibody Characterization. Antigen, Immunogen, Manufacturer, Dilution factor. Cholera toxin subunit B, Purified CTB isolated from Vibrio cholerae, List Biological Laboratories (Campbell, CA), goat polyclonal #703, 1:5,000. …

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