Bacterial Toxin Research Citations

Anthrax toxin neutralization assay (TNA):

The anthrax TNA was performed using the mouse macrophage cell line J774A.1 (European collection of cell cultures, ECACC) as previously described [25] with slight modifications. 96-well flat-bottomed tissue culture plates (Greiner Bio One, Germany) containing 810⁴ macrophages/well in DMEM (Life Technologies, USA) and 10% FCS (Life Technologies, USA) were incubated overnight at 37C and 5% CO₂. Goat sera were doubly diluted (1:50 to 1:6400) in culture medium containing PA and LF (List Biological Laboratories Inc., Campbell, CA) at concentration of 500 ng/ml and 400 ng/ml (lethal toxin, LT) respectively. The sera/LT mixture was incubated for one hour at 37C and 5% CO₂ before adding to overnight cultured cells (after discarding medium) and incubated for three hours. …

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

Adoptive transfer experiments:

Enriched CD4⁺CD25⁺ cells were then cultured in complete medium alone or with pertussis toxin (ptx; List Biological Laboratories) at 100 ng/ml for 2 h in vitro and transferred to congenically marked recipient mice. Splenocytes of recipient mice were harvested 36 h after transfer and analyzed by flow cytometry.

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)

Materials:

B. anthracis PA protein was from List Biological Laboratories, Inc. (Campbell, CA).

Adsorption of protein antigens on aluminum hydroxide particles:

The adsorption of proteins (OVA or PA) on aluminum hydroxide particles was carried out by mixing the particles in suspension with the protein in solution. Briefly, a certain volume of the protein solution was added into a tube (10 g OVA or 4 g PA), followed by the addition of particles in suspension at a weight ratio of 1:5 to 1:1 (OVA vs. particles) or 1:5 (PA vs. particles). After 20 min of gentle stirring, the protein-particle mixtures were stored at 4C or freeze-dried, if needed, before further use.

Immunization studies:

…When PA was used as the antigen, female BALB/c mice (1820 g) were immunized subcutaneously with PA-adsorbed aluminum hydroxide particles on days 0 and 14. As negative controls, mice were injected with sterile PBS or PA alone. The dose of PA was 4 g per mouse per injection, and the dose of the particles was 20 g per mouse per injection.

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

Reagents:

cholera toxin (List Biological Laboratories, Campbell, CA, USA)…

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

Materials:

…Lipopolysaccharide (LPS)…were purchased from List Biological Laboratories, INC. (CA, USA)

In vitro phototoxicity testing:

Raw 264.7 macrophage cells were seeded in a 96-well plate at 1 10⁴ cells/well, incubated for 24 h for cell attachment. Activated macrophages were obtained by treating Raw 264.7 cells with LPS (1 g/mL) and INF- (50 ng/mL) for 24 h.

Author did not specify which List Labs LPS product was utilized in their research.  List Labs provides the following LPS products:  https://listlabs.com/product-information/lipopolysaccharides/

In Vitro CD4+ T-Cell Polarization Assays:

DC subsets sorted from spleens, mLN, or cLP of mice were incubated with endotoxin-free OVA (1 mg/mL; BioVendor) and/or 0400 ng/mL LPS (List Biological Laboratories) before coculturing with OT-II nave CD4+ T cells (1:10 ratio) in DMEM…

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

…For peptide immunization of Balb/c mice, 100 µg of peptide was mixed with 5 µg of holotoxin CT (List Biological Laboratories, Campell, CA) in a total of 20 µl of PBS and 10 µl was administered in to each nare. …

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

Labeling of Retinal Projections:

Six animals (three O. degus and three O. lunatus) were anesthetized as previously mentioned, and given intraocular injections (12 L, left eye) of 0.5% Cholera toxin subunit B (CTB, List Biological Laboratories) mixed with 1% DMSO.

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

In vitro cell stimulations and in vivo treatments:

For PTX treatment experiments, pDC or CD11b⁺ cells were purified from 2 – 3 pooled spleens, then preincubated for 30 min with 100 ng/ml pertussis toxin from B. pertussis (List Biological Laboratories, Inc.) prior to stimulation.

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)

Reagents and Materials:

… Protective antigen (PA) protein of Bacillus anthracis was purchased from List Biological Laboratories, Inc. (Campbell, CA; USA). …

Rapid Peptide Specificity Analysis:

The peptide specificity analysis was performed by CE using PA (positive control), BSA, HRP, and an anti-PA mAb at 20 kV as described above. …

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