Bacterial Toxin Research Citations

Reagents:

Cholera toxin (CT) was purchased from List Biological Laboratories Inc. (Campbell, CA, USA). …

Experimental design: oral sensitization, immunotherapy and challenges:

Mice were sensitized intragastrically (i.g.) to whey (20 mg in 0.5 ml PBS) or PE (6 mg in 200 l PBS) using CT (15 µg/mouse) as an adjuvant [23, 24] (day 0, 7, 14, 21 and 28 for whey and day 0, 1, 2, 7, 14, 21 and 28 for PE) (Fig. 1). Sham-sensitized mice received CT in PBS alone. …

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

… LPS from E. coli O111:B4 was obtained from List Biological Laboratories, Inc. (Campbell, CA) and resuspended in 50 mM NaPO4, 300 mM NaCl, pH 7 buffer at its solubility limit of 200 M. …

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

… Goat anti-CTb primary antibody List Biological Laboratories CTb 703; …

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

Pertussis toxin List Biological Laboratories

Cellular assays – PTX, TY52156 treatment of BP:

For treating BP with pertussis toxin (PTX) or S1PR3 inhibitor TY52156, pericytes were seeded in 6 wells (100 000 pericytes) or 6 cm dishes (400 000 cells). Pericytes were treated with 1 M inhibitor (dissolved in 100 % ethanol) or 200 ng of PTX (dissolved in PBS) with corresponding controls (100 % ethanol or PBS) for 4 hours in PM containing 0.5 % FCS without growth factors. Subsequently, cells were stimulated with S1P and harvested for further analysis.

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)

EAE Induction:

… On the day of immunization and 2 days after, mice also were injected intravenously with 200 ng pertussis toxin (List Biological Laboratories, Campbell, CA, USA). … After culture for 3 days, cells were collected and washed in PBS. 1x 10⁷ splenocytes were transferred into Rag2^/ mice by intravenous injection. Recipient mice also were given 200 ng pertussis toxin intravenously on the day of cell transfer and 2 days after transfer. …

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)

EAE induction:

EAE was induced by s.c. immunization of mice at the flanks with an emulsion of MOG₃₅₅₅ peptide (100g) emulsified in CFA supplemented with 4mg/ml Mycobacterium tuberculosis extract H37Ra (Difco). Additionally, the animals received 200 ng pertussis toxin (List Biological Laboratories) i.p. on days 0 and 2. Clinical signs of EAE were assessed daily and scored according to the following criteria: 0, no signs of disease; 1, limp tail; 2, hind limb weakness; 3, hind limb paralysis; 4, hind limb and forelimb paralysis.

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)

… 25 l of serial 1:3 dilutions of rabbit sera were mixed with 50 l of Stx1 (List Biological Laboratories, Campbell, CA, USA) in a concentration of 4 CD50/ml in wells of a separate plate, incubated at 37o C in 5% CO2 for one hour, …

• Product #161 – Shiga Toxin 1 from Escherichia coli

Dorsal hemisection SCI with sciatic nerve conditioning injury:

Wildtype C3^+/+ mice used in this study were litter-mates of C3^/ mice. … Five days prior to tissue collection, the left sciatic nerve was exposed again and 1.5 µl of 10 mg/mL cholera toxin subunit (CTB, List Biological Laboratories), a trans-ganglionic tracer, was injected into the proximal nerve section using a pulled glass capillary tube pipette. …

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

Experimental schedule:

Cells were stimulated with LPS (0.2 g/mL; List Biological Labo., Campbell, CA, USA) or nigericin … The expression of inflammasome-related molecules and the level of secreted IL-1 were analyzed by quantitative RT-PCR/western blotting and an enzyme-linked immunosorbent assay (ELISA), after culture for 6 h and 24 h, respectively. Trophoblasts were incubated with N6, 2 -0- dibutyryladenosine 3, 5-cyclic monophosphate (db-cAMP; 0.5 mM; SigmaAldrich) to enhance their fusion toward syncytiotrophoblasts in DMEM supplemented with 2% FBS for 2 days.

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/

Assessment of light chain protease activity (VAMP-2):

The protease activities of BoNT/B1 (List Laboratories, US), rBoNT/B1 and rBoNT/B1_(S201P) against VAMP-2 were assessed using the BoTest (Biosentinel, Wisconsin, US) cell-free assay. BoNT/B1, rBoNT/B1 and rBoNT/B1_(S201P) were diluted to 1.39 nM in BoTest Reaction Buffer (50 mM HEPES-NaOH, 5 mM NaCl, 10 M ZnCl₂, 0.1% Tween-20, 0.1 mg/mL BSA, pH 7.1). BoNTs were reduced at room temperature (20 2C) for 30 minutes by addition of 5 mM DTT to allow maximum catalytic activity in the assay. BoTest Reporter (VAMP-2 (3394) flanked by N-terminal cyan fluorescent protein (CFP) and C-terminal yellow fluorescent protein (YFP) in 50 mM HEPES-NaOH, 10 mM NaCl, 15% glycerol) at a final concentration of 200 nM was combined with reduced BoNT/B1, rBoNT/B1 or rBoNT/B1_(S201P) (final concentration 0.5 pM1.25 nM) in black Maxisorp plates (Nunc) in a final assay volume of 100 µL/well. The plates were sealed, wrapped in aluminium foil to prevent degradation of the light-sensitive substrate and incubated at 30°C for 18 h. After incubation, fluorescence emission at 485/20 nm and 528/20 nm following excitation at 440/40 nm was measured using a BioTek Synergy HT plate reader.