Bacterial Toxin Research Citations

EAE induction and activity

Ten- to 12-week-old mice were injected subcutaneously with 200 μg of MOG35-55 peptide [GL Biochem (Shanghai) Ltd.] in Freund’s complete adjuvant containing heat-killed Mycobacterium tuberculosis H37Ra (5 mg/ml; BD Difco). On the day of immunization and 48 hours later, 200 ng of pertussis toxin (List Biological Laboratories) was administered intravenously.

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)

… pertussis toxin was purchased from List Biological Laboratories (Campbell, California, USA), …

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 were anesthetized with isoflurane and injected with two 100 µL emulsion injections along the midline. Two and 24 h later, mice were injected intraperitoneally (i.p.) with 60 ng of PTX (List Biological Laboratories). In male mouse studies, 40 ng of PTX was used. Clinical signs of disease were monitored accordingly: …

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)

STAR★Methods
Key resources table

… Diphteria toxin, List biological Labs …

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

Experiment 1 (Figure 1): Muscle reinnervation after blocking synapses with Tetanus toxin (TeTx). In this experiment the animals underwent a muscle injection in the left tibialis anterior (TA) muscle of either TeTx or vehicle (bacteriostatic sterile saline, Med-Vet International, Mettawa IL, USA). TeTx (Lot#19050A1) was obtained from List Biologicals (Campbell, CA, USA). …

• Product #190 – Tetanus Toxin from Clostridium tetani

Pyroptosis inducers and reagents

For activation of the NLRC4 inflammasome, late-log cultures of Salmonella enterica serovar Typhimurium strain SL1344 (a kind gift from Dr. Brad T. Cookson, University of Washington) were grown in Luria-Bertani broth containing 0.3 M NaCl with shaking at 37 °C. Bacteria were washed, resuspended in PBS, and added at a multiplicity of infection of 10 bacteria per macrophage for 90 minutes (unless otherwise indicated). For NLRP1b activation, BMDM from BALB/cJ mice were treated for two hours (unless otherwise indicated) with 1 µg/mL anthrax lethal toxin, prepared as a 1:1 mix of protective antigen and lethal factor (List Biological Labs). For NLRP3 activation, BMDM from C57BL/6 J mice were primed with 100 ng/mL of LPS (List Biological Labs) for three hours, followed by addition of 15 µM nigericin (Cayman Chemicals) for an additional three hours. For pyrin activation, BMDM from C57BL/6 J mice were primed with 500 ng/mL of LPS (List Biological Labs) for three hours, followed by addition of Toxin A (TcdA) from Clostridium difficile (2 µg/mL) (List Biological Labs). For AIM2 activation, BMDM from C57BL/6 J mice were primed with 500 ng/mL of LPS (List Biological Labs), before transfection of 2 µg/mL poly (dA:dT) (Invivogen) at a 1:25 ratio of DNA to lipofectamine (µg/µL) utilizing Lipofectamine 2000 + OptiMEM, as described [53]. Thiomuscimol (Santa Cruz Biotechnology and Cayman Chemicals) and muscimol hydrobromide (Sigma Aldrich) were added at specified concentrations, at the same time as the pyroptosis inducer, unless otherwise specified. The PDI inhibitors PACMA31 and CCF642 (Cayman Chemicals) were reconstituted in 100% DMSO and stored as frozen aliquots at −20°C before use. Each were diluted to target concentrations of 5 µM for CCF642 and 2.25 µM for PACMA31 in cell culture medium, resulting in a final DMSO concentration of 1%.

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

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

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

• Product #152C – Toxin A from Clostridium difficile

… To produce foam cells, BMMs were treated with 100 µg/mL of acLDL and 100 ng/mL of LPS (List Biological Labs Inc, Cat. 314), simultaneously, for 24h (RNA) or 48h (protein and other experiments) …

• Product #314 – LPS from Escherichia coli K12 strain LCD25

2.5. Surgery

… Epileptic rats receive TeNT (25 ng in 250 nl, or 75 ng in 750 nl following a decrease in toxicity due to a freezer malfunction; List Biological Inc., Campbell CA, supplied by Quadratech Diagnostics, Lewes, UK). Batches of toxin can vary in their toxicity, so we routinely assay them (Mellanby et al., 1968; Barkmeier and Loeb, 2009; Sunderam et al., 2009; Wykes et al., 2012). …

• Product #190 – Tetanus Toxin from Clostridium tetani

Animal experiments

… For the oral immunization model, the mice received sodium bicarbonate solution to neutralize stomach acid and were orally immunized with 1 mg of OVA (Sigma) or 1 mg of NP-OVA (Biosearch Technologies) plus 10 μg of CT (List Biological Laboratories) on days 0, 7 and 14. One week after the final immunization, fecal samples and mononuclear cells from the intestinal lamina propria (iLP) were collected for enumeration of OVA- or CTB-specific antibody responses via ELISA and ELISPOT. …

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

… DT (List Labs, cat. no. 150) was dissolved in 1× PBS and administered by intraperitoneal injection …

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