Bacterial Toxin Research Citations

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

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

Toxin Neutralization Assay (TNA):

… Goat sera (starting dilution; 1:50) were doubly diluted in culture medium containing PA and LF (List Biological Laboratories, Inc …

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

Chemicals and reagents:

All bacterial toxins were purchased from List Biological Laboratories (Campbell, CA). …

Human B-lymphocytes sensitivity to Pseudomonas toxin-mediated lethality:

Human B lymphocytes were treated with serial dilutions of P. aeruginosa exotoxin A for 48hours.

Cellular drug screens:

RAW264.7 cells (10,000 per well) were seeded in 96-well plates 24hours before the assay. Cells were treated with compounds for 1hour, and then challenged with either 2g/ml PE or 4g/ml cholera toxin for 12hours. As rodent cells are insensitive to diphtheria toxin, C32 cells were treated with 2g/ml of diphtheria toxins for 24hours. …

Toxins treatments and cell viability assays:

Cells (10,000 per well) were seeded in 96-well plates 24hours before the assay. Cells were treated with Bithionol for 1hour. RAW264.7 cells were challenged with anthrax toxins that include LF or FP59 and PA83 or PA63 (for 6hours), PE (for 12hours), or cholera toxin (for 12hours) at 0.5, 2, and 4g/ml respectively. C32 cells were treated with 2g/ml of diphtheria toxin for 24hours. Determination of cells viability was performed by MTT assay….

MAPKK2 cleavage assay:

… Following pre-treatment, the cells were exposed to 0.5g/ml of PA and LF at 37C for up to three hours in the presence of 33M of Bithionol. …

Cellular cathepsin B and caspase-1 activity assays:

…Caspase-1 activities were induced by 1hour pretreatment of cells with 0.5g/ml of LF+PA, and was compared to cells un-induced by the toxin. …

BoNT/A light chain FRET assay:

The reaction volume was 250l per well in 96 wells plate, containing 50mM HEPES pH 7.4 containing 0.05% TWEEN 20, 5M SNAPtide (BoNT/A substrate peptide) conjugated with DABCYL and FITC (List Biological Laboratories, Inc), and 33M of Bithionol. …

• Product #160 – Exotoxin A from Pseudomonas aeruginosa
• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• 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
• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis
• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin
• Author did not specify which Cholera toxin was utilized.  List Labs provides Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae and Product #101 – Cholera Toxin from Vibrio cholerae.

…Recombinant protective antigen (rPA) was purchased from List Laboratories. …

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

Reagents:

Anthrax lethal toxin, consisting of protective antigen and lethal factor was 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

IL-1 signaling assay:

… or Bacillus anthracis lethal factor (100 ng/ml; List Biological Laboratories) was prepared with these liposomes as controls. …

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).

Materials: 

…The Anthrax PA measured by circular dichroism was obtained from List Labs (Lot: 17115A6B). …

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

Western Blot Assays:

…Primary antibody was then added in 10 ml of 5% NFDM in TBST…and for PA, anti-protective antigen from B. anthracis, goat, 1:5,000, List Biological Laboratories, Campbell, CA) and incubated overnight at 4C. …

…ELISAs were performed as described [123]. Briefly, 96-well plates were coated with 1 g/well of recombinant PA, LF (List Biologicals, Campbell, CA), …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Author did not specify which List Labs Lethal Factor was utilized.  List Labs offers Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).