Bacterial Toxin Research Citations

3.2. Reagents

We obtained purified lipopolysaccharides (LPS) from Escherichia coli from List Bio-logical Laboratory (Campbell, CA, USA), which we dissolved in endotoxin-free water. …

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/

Chemical reagents

… Cholera toxins from Vibrio cholerae (product no.100B: Lot no. 10071A2) were purchased from List Biological Laboratories (Campbell, CA, USA). …

Human colonoid swelling assays

Human epithelial crypt-like cells were obtained from colon tissue biopsies. Crypt isolation was performed as previously described with minor modifications [23]. … Swelling of the human colonoids was monitored by random imaging every 5 min using the BioTek Cytation™ 5 Cell Imaging Multi-Mode Reader after treatment with forskolin (5 μM), forskolin plus nornidulin, cholera toxin (2 μg/mL), or cholera toxin plus nornidulin. Swelling of colonoids was calculated as the relative area increased from 0 min to 60/240 min and analyzed as the area under the curve (AUC) using CellProfiler 4.2.1 program.

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

Anthrax lethal toxin neutralization assay

RAW264.7 cells were seeded as mentioned above and cultured in DMEM supplemented with 10% FBS and 10 mM HEPES (Fisher Scientific, Fai Lawn, NJ, USA) overnight. The toxin neutralization assay was performed as previously described [73]. Pooled serum samples from vaccinated guinea pigs before challenging with Bcbva UFBc0009 and B. anthracis Ames UF00738 spores were prepared at 200-, 1,200-, and 12,000-fold dilutions in DMEM supplemented with 10% FBS and 10 mM HEPES. Pooled serum samples from unvaccinated animals served as a control. The diluted serum samples were then incubated with 50 ng/ml PA (List Labs, Campbell, CA, USA) and 160 ng/ml LF (List Labs, Campbell, CA, USA) for 30 min before adding to RAW264.7 cells. …

• 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

Mouse treatments. Mice received 3 doses of i.p. tamoxifen (Sigma Pharmaceuticals) over one week. Tamoxifen was suspended in corn oil and delivered at a concentration of 0.2 mg/g per dose. Animals were rested for 4 weeks after the final tamoxifen dose (13, 38). E. coli LPS O55:B5 (List Biological Laboratories) was instilled into the tracheas of mice sedated with isoflurane (Baxter) at a concentration of 20 μg in 50 μL of PBS using a modified gel-loading tip with direct laryngoscopy (13). Bleomycin (TEVA Pharmaceuticals) was administered at a dose of 1.5 U/kg in 50 μL of PBS by the same method (21). Mice were euthanized by i.p. injection of 10% Fatal Plus solution (Vortech Pharmaceuticals). Three to 4 mice were treated per time point in each inflammatory model.

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

… Eight days after oral administration of OVA, 1 mg OVA + 20 µg cholera toxin (100B; List Biological) in 0.2 M sodium bicarbonate were provided by gavage once per week …

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

Animals

… Male, Advillin-Cre+/−, mice were crossed with female, CalcaWT/GFP (Figure 1D), mice to generate CalcaWT/GFP, CalcaWT/DTR (Figure 1C), or wildtype CalcaWT/WT (Figure 1A) offspring. Diphtheria toxin (DTX; List Biological Laboratories, Product #150) was reconstituted to 20 μg/mL in sterile 0.9% saline, aliquoted, and stored at −80°C until used for injections. Once animals reached 10–12 months of age, DTX was administered by i.p. injection (100 μg/kg) twice with 72 h between the two injections. Animals were sacrificed and prostate tissue was harvested 14 days after the second injection (Figure 1E).

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

… Mice were subcutaneously injected with 100 µg of the emulsion under each armpit, and Pertussis Toxin (PTX, 200 ng/mouse, List Biological Laboratories, 180, USA) solution was injected into the tail vein on the day of immunization (day 0) and the following day after …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

2.3 Study design and induction of EAE and assessment of EAE severity

… Experimental autoimmune encephalomyelitis (EAE) was induced on day 28 and developed until day 46. Mice were immunized by two subcutaneous injections on both flanks, each containing 50 µg of myelin oligodendrocyte glycoprotein peptide MOG35-55 (ProSpec-Tany Techno Gene, Ness-Ziona, Israel) emulsified in 50 µL complete Freund’s adjuvant, supplemented with 5 mg/mL of heat-inactivated Mycobacterium tuberculosis H37Ra (Sigma-Aldrich). Mice also received two doses of 300 ng pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA) intraperitoneally on the day of immunization and 24 hours later (23). …

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 from Bordetella pertussis from List Biological Laboratories, …

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)

2.2. EAE induction and evaluation

Each mouse was injected subcutaneously with a 0.2 mL emulsifier containing 50 % MOG35–55 (Genescript, USA) and 50 % complete Freund’s adjuvant (CFA; Sigmasingle bondAldrich, USA) at the posterior neck and root of the tail on day 0. The emulsifier was prepared from 200 µg of MOG35–55 dissolved in 0.1 mL of PBS with an additional 4 mg/mL Mycobacterium tuberculosis H37Ra (Sigmasingle bondAldrich, USA). At the same time, 300 ng of Pertussis toxin (PTX) (List Biological Laboratories 181, USA) was injected intraperitoneally on days 0 and 2. Clinical signs of EAE were scored as follows (Stromnes and Goverman, 2006): …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)