Bacterial Toxin Research Citations

Acute lung inflammation model:

LPS (E. coli O55:B5 from List Biological Laboratories) (20 g) in 50 l of PBS was instilled directly into the tracheas of mice sedated with isofluorane (Baxter) using a modified feeding needle (59). Mice were sacrificed using i.p. injection of pentobarbital sodium solution.

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

… B. anthracis LF was purchased from List Biological Laboratories. …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Product #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

EAE Induction:

… In addition, 500 ng pertussis toxin (PT; List Biological Laboratories, CA, USA) was injected by intraperitoneally on days 0 and 2 after immunization. …

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)

Chemicals and Reagents:

Diphtheria toxin (DT) was obtained from List Biological Laboratories (Campbell, CA. Cat. No. 150), reconstituted in PBS and stored at 80C. Mice received a dose of DT (10ngg¹ in 0.1ml PBS)³⁰, injected intraperitoneal, every other day.

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

Reagents:

… B. anthracis lethal factor (LF) was acquired from List Biologicals (#172C). …

J774.A1 culture and stimulation:

J774.A1 cells were cultured in IMDM (Lonza) containing 10% (vol/vol) heat-inactivated FBS, 1% (vol/vol) nonessential amino acids (Lonza), 100 U/ml penicillin, and 100 mg/ml streptomycin at 37C in a humidified atmosphere containing 5% CO₂. Cells were scraped for passages and, for experiments, counted and seeded into 96-well plates in culturing medium. On the next day, cells were changed to fresh media and stimulated with vehicle or 1 g/ml PA combined with 0.5 g/ml LF (LeTx), VBP (0.1% TFA in DMSO) or 1G244 (DMSO) at a final concentration of 10 M or 25 M.

• Product #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

… To compare the potency of BoNT/D with BoNT/A (subtype BoNT/A1; List Laboratories Inc.), the drug substance of incobotulinumtoxinA, concentration-response curves were constructed from three data points, each determined at least in triplicate, fitted to the logarithmic functions y(BoNT/D) = -25.548Ln(x) + 233.4, R2 = 1.0 and y(BoNT/A) = -37.817Ln(x) + 259.88, R2 = 0.9885. …

EXPERIMENTAL –  FRET Lethal Factor Protease Assay:

The FRET assay measures the enzymatic activity of LF with an internally quenched peptide substrate derived from MAPKK. LF cleavage of the substrate separates the quencher and fluorescent tag leading to an increase in fluorescence intensity. The slope of fluorescence absorbance is therefore used to determine enzyme activity. Anthrax lethal factor (30 nM final concentration) and MAPKKide substrate (25 M final concentration) were purchased from List Biological Laboratories, Campbell, CA. …

Author did not specify which MAPKKide substrate was utilized; List Labs provides the following:

• Product #530 – MAPKKide Peptide Substrate (o-Abz/Dnp) for Anthrax Lethal Factor
• Product #531 – MAPKKide Peptide Substrate (DABCYL/FITC) for Anthrax Lethal Factor
• Product #532 – MAPKKide Plus (AMC) Specific Substrate for Anthrax Lethal Factor

K⁺ bead assays:

… For intraphagosomal K⁺ bead assays, primary murine bone marrow-derived macrophages (2×10⁵/well) were seeded in a 96-well clear bottom black plate. Where needed, 100 U/ml IFN (PeproTech) and 10 ng/ml LPS (List Biological Laboratories) were used for activation of macrophages. 100 nM concanamycin A (AdipoGen Life Sciences) was added to the media where indicated. Macrophages were washed three times with pre-warmed assay buffer, before addition of APG4/AF594 beads (~25 beads/macrophage). A Biotek Synergy H1 microplate reader was used for fluorescence tracking, with four-six replicate wells/condition and bottom reads taken every two minutes for four hours. Temperature was maintained at 37C throughout the assay.

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/

Animals and induction of EAE:

… Mice in the EAE group were treated with 1 mg/ml of the MOG_35-55 peptide (purity >96.44%; GL Biochem Ltd., Shanghai, China) emulsified in complete Freund’s adjuvant (Difco Laboratories Inc., Detroit, MI, USA) and supplemented with 5 mg/ml Mycobacterium tuberculosis H37Ra (Difco Laboratories Inc.) by subcutaneous injection into the hind flank. Pertussis toxin (500 ng; List Biological Laboratories, Inc., Campbell, CA, USA) was injected intraperitoneally at days 0 and 2 post-immunization (PI) …

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)

Mice, PCR, and EAE immunization:

To induce EAE, adult female mice were injected subcutaneously in the flank and at the tail base with 200µg of myelin oligodendrocyte glycoprotein (MOG) 3555 peptide emulsified in complete Freunds adjuvant (BD Biosciences, San Jose, CA, USA) supplemented with 600 g of Mycobacterium tuberculosis (strain H37Ra; BD Biosciences). Two i.p. injections of 400 ng pertussis toxin (List Biological Laboratories, Denver, CO, USA) were given 0 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)