Bacterial Toxin Research Citations

Antibodies and reagents
… mutant pertussis toxin was purchased from List Labs (cat no. 184). Purified cholera toxin was purchased from List Labs (cat no. 100B). …

• Product #184 – Pertussis Toxin Mutant
• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

2.1. Animals and EAE induction

Eight-week-old female C57BL/6N mice were obtained from the University of Seville Animal Facility and housed at the IBiS Animal Facility under a 12 h light/dark schedule (lights on at 8:00 a.m.) and ad libitum access to water and food. EAE was induced in the animals by subcutaneous immunization in both hind legs with 100 μg of MOG35–55 (Cambridge Research Biochemicals, Cleveland) emulsified in CFA (Sigma) containing 50 μg of heat-killed Mycobacterium tuberculosis (H37Ra, ATCC 25177) and two doses of intraperitoneal pertussis toxin (200 ng/day) (List Labs, California) on days 0 and 2 post-induction. …

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)

Induction and evaluation of experimental autoimmune encephalomyelitis (EAE)

For EAE experiment, each mouse was injected with 50 μg myeline oligodendrocyte glycoprotein (MOG35-55: MEV GWY RSP FSRVVH LYR NGK; GenScript) emulsified in incomplete Freund’s adjuvant (Chondrex, Inc.) and 50 μg desiccated Mycobacterium tuberculosis (strain H37RA). On the same day of immunization and 2 days after, mice received 300 ng/mouse pertussis toxin (List Biological Laboratories) via intravenous injection. …

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)

Ex Vivo Cultures

Biopsy tissues (5 mm2) were cultured in triplicate in 12-well plates containing a modified Connaught Medical Research Laboratory–based cell-culture medium. Tissues were positioned such that the apical mucosal surfaces faced the liquid-air interface. Culture plates were incubated in medium (37°C, 5% CO2) with either test compounds or vehicle. Tissues were treated with Staphylococcus enterotoxin B (List Biological Laboratories, Campbell, CA) to stimulate inflammatory cytokine secretion. Cell-culture supernatants (approximately 1 mL) were collected and flash frozen 18 hours after treatment. Two samples from each condition were stabilized in RNAlater (Invitrogen, Carlsbad, CA) and frozen.

2.3. Active EAE induction and treatments

The EAE model was induced in female C57BL/6 mice using active induction as previously described [40], [41]. Briefly, we prepared a uniform emulsion containing 200 µg of myelin oligodendrocyte glycoprotein (MOG35–55, GenScript, Nanjing, China) mixed with an equal volume of complete Freund’s adjuvant (CFA, Sigma, US) containing 5 mg/mL of Mycobacterium tuberculosis H37Ra (231141 Dessicated, BD, Difco, US). This emulsion was subcutaneously injected (s.c.) into the neck and back of the mice on day 0. Subsequently, a total of 200 ng of pertussis toxin (PT, List Biologic, Campbell, CA, US) was administered intraperitoneally (i.p.) to the mice on days 0 and 2 following 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

… LF (List Labs), …

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

Experimental EAE

Female age-matched Smarcd2fl/fl and Cd4creSmarcd2fl/fl mice, Foxp3cre−YFP and Foxp3cre−YFPSmarcd2fl/fl mice (8–10 weeks old) were immunized subcutaneously (s.c.) with 200 μg of MOG35–55 peptide (MEVGWYRSPFSRVVHLYRNGK, ChinaPeptides) emulsified in complete Freund’s adjuvant (CFA) (Sigma, F5506) containing heat-killed Mycobacterium tuberculosis (BD, 231143). In addition, the mice were administrated 200 ng of Pertussis toxin (List Biological Laboratories, 181) intravenously (i.v.) on days 0 and 2. …

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

… DTX List Biologicals …

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

… Botox injection to induce paralysis of whisking43,85-87. Both whisker pads were injected with Botox (Botulinum Neurotoxin Type A from Clostridium botulinum, List Labs #130B). A stock solution of 40 ng/µl …

Immunocytochemistry

After post-fixation, brains were submerged in 10% sucrose PBS solution at 4□ until they sank. The brains were then embedded in 14% gelatin and 10% sucrose, after which they were fixated in 10% formalin and 30% sucrose overnight, and subsequently in 30% sucrose until they sank. The brains were then cut in 50 μm sections. For light-microscopic staining of the CTb, the sections were first incubated for 20 mins in 3% H2O2 (in PBS) to remove endogenous peroxidase activity of blood. Next, the sections were blocked in 10% normal horse serum (NHS) in PBS with 0.5% triton, after which they were incubated at room temperature with anti-CTb (goat, 1:15000, List labs) primary antibody (in PBS with 2% NHS, 0.5% triton) for 48-72 hrs and next with biotinylated rabbit anti-goat (1:200, Vector) secondary antibody (also in PBS with 2% NHS and 0.5% triton) for 1.5 hrs. …

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