Bacterial Toxin Research Citations

Primary Cells Extracted from Human Corneal Biopsies

Td_hLESCs were collected from scleral‐corneal tissue (obtained with written consent from next of kin according to the directives set by the Italian Centro Regionale Trapianti and Centro Nazionale Trapianti, and only when not suitable for transplantation) and preserved in media before processing. Biopsies, composed of cornea, limbus, and conjunctiva, were dissected into quarters, and then the cornea and conjunctiva were removed. The limbus was fragmented and soaked in PBS. Trypsin/EDTA (0.05%/0.01% w/v; Thermo Fisher Scientific, Italy) was added (10 mL) to digest the tissue for 30 min at 37 °C. The solution was centrifuged for 5 min at 1000 rpm and the pellet resuspended in fresh media. This cycle was repeated thrice to extract the majority of the cells. Cells were plated into a 24‐well plate (30 000 cells per well) in coculture with irradiated 3T3‐J2 cells. Culture medium consisted of Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F12 (DMEM/F12, 2:1, Thermo Fisher Scientific) supplemented with 10% v/v FBS (Thermo Fisher Scientific), 50 µg mL–1 penicillin‐streptomycin (P/S), 4 × 10−3 m glutamine (EuroClone, Italy), 5 µg mL–1 insulin (HUMULIN R, Lilly, Canada), 0.4 µg mL–1 hydrocortisone (Flebocortid Richter, Sanofi, Italy), 0.18 × 10−3 m adenine (adenine grade I, Pharma Waldhof GmbH, Germany), 8.1 µg mL–1 cholera toxin (Cholera Toxin QD, List Biological Laboratories, USA), 2 × 10−3 m triiodothyronine (Liotir, IBSA, Italy), and 10 ng mL–1 epidermal growth factor (GMP Cellgro, CellGenix GmbH, Germany).

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

… For macrophage activation, the adherent cells were stimulated with 1g/mL of lipopolysaccharide from E.coli O55:BS (List Biological Lab Inc.) in media for 4 h at 37 C …

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

… Retrograde tracing was performed using a 1% solution of cholera toxin B subunit (CTB; List Biological Labs), injected using standard stereotaxic coordinates (Paxinos and Franklin, 2001). …

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

… we iontophoresed CTb (0.1% low- salt CTb in distilled water; #104, List Biological) through a glass micropipette with the tip NTS sections for CTb with a goat polyclonal antiserum (1:10,000; List Biological, 703 …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

Reagents

Purified lipopolysaccharides (LPS) from Escherichia coli were obtained from List Biological Laboratory (Campbell, CA, USA) and dissolved in endotoxin-free water. …

Analysis of Inflammasome Activation

This was performed as described previously [33]. BMDMs were seeded at 2 × 106 cells/well in 6-well plates for immunoblot assay and 4 × 105 cells/well in 96-well plates for ELISA. BMDMs were primed with LPS for 4 h. To exclude the effect of sweroside on the LPS priming step, sweroside was added after washing out the LPS with phosphate-buffered saline. …

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/

… Intravenous LPS challenge. Intravenous LPS challenges were only performed on occasion 2 and 3. Subjects received 1 ng/kg (cohort 1) or 2 ng/kg (cohort 2) E. Coli-purified LPS (GMP-grade from Lot#94332B4, List Biological Laboratories Inc …

• Product #9433 – GMP LPS Lipopolysaccharide List™ Hpt™ From Escherichia Coli Type O113

Antibody quantification by ELISA

PA, LF and EF ELISAs were performed using commercially available antigens (List Biologicals, Campbell, CA) as previously described [21]. Cutaneous anthrax and regional control group sera were titered using 2-fold dilutions, beginning at 1:10. US control sera were titered at 10-fold dilutions, beginning at 1:10. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #178A – Anthrax Edema Factor (EF), Recombinant from B. anthracis

… A microelectrode filled with a retrograde tracer, 1% CTb (List Biological Laboratories, Campbell, CA, USA) dissolved in 0.1 M phosphate buffer or 1% FG (Fluorochrome, Englewood, CO, USA) dissolved in saline was used. CTb or FG was electrophoretically (for 510 min with 2.0-A positive, 300 ms duration pulses at 2 Hz) injected into the OPC identified electrophysiologically as in the second experiment …

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

… Sections were incubated in a primary antibody against the inhibitory neuron transcription factor Pax2 (1:500 dilution, ThermoFisher #716000; RRID: AB2533990) or in antibodies against CTB (1:2000 dilution; List Biological Catalog #703, RRID: AB_10013220 …

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

Materials and Methods

A total of 40 female C57BL/6 mice, 10-week-old, were randomly divided into four groups, including induced control groups without EAE induction, induced EAE without training, and forced and voluntary training groups. All animals have started training 4 weeks before experimental autoimmune encephalitis- (EAE-) induction. The mice were intraperitoneally anesthetized with an injection of xylazine (5 mg/kg) and ketamine (50 mg/kg). Myelin oligodendrocyte glycoprotein peptide 35–55 (300 μg in saline; MOG 35–55; KJ Ross-Petersen ApS, Denmark) was emulsified in an equal volume of complete Freund’s adjuvant (CFA; Sigma, USA) containing 500 μg of heatkilled Mycobacterium tuberculosis. This emulsion was then injected subcutaneously at the base of the tail of each mouse. The mice also received intraperitoneal injection of pertussis toxin (300 ng in 10 μl PBS; List Biological Lab, USA) at the time of immunization and the same was repeated 48 hours later. To confirm EAE induction, animal clinical assessment of EAE was performed daily using a 15-point scoring system following immunization as reported previously by Giuliani et al. [13].

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)