Bacterial Toxin Research Citations

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

Experimental Autoimmune Encephalomyelitis

For MOG35–55 induced EAE, age (8–12 weeks old) and gender matched mice were subcutaneously immunized (s.c.) with 50 μg of MOG35–55 peptide (MEVGWYRSPFSR VVHLYRNGK, AnaSpec) and 500 μg of M. tuberculosis (Difco) emulsified in IFA (Difco). In addition, the mice were administrated 200 ng of Pertussis Toxin (List Biological Laboratories) intraperitoneally (i.p.) on days 0 and 2, respectively. For adoptive transferred EAE, 2D2 CD4+ T cells were differentiated under pathogenic Th17 differentiation conditions for 4 days and re-stimulated in fresh medium in the presence of plate-bound anti-CD3 (2 µg/ml) and anti-CD28 (2 µg/ml) antibodies for 2 days. Live cells were purified as described. Approximately 1 × 106 purified cells were i.v. injected into Rag1−/− mice. Approximately 200 ng of Pertussis toxin (List Biological) was injected intra-peritoneal (i.p.) on days 0 and 2 after T cell transfer. For induced SKI expression in adoptive T cell transfer induced EAE, tamoxifen (TMA group) or corn oil (Control group) were injected intraperitoneally at days 1 and 3 after EAE elicitation as illustrated. …

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)

Protective Antigen List Biological Labs #171E

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

Pertussis toxin, List Biological Laboratories, Cat# 181

EAE induction

To induce EAE, a 1:1 incomplete Freund’s adjuvant (IFA) to PBS emulsion was prepared with final concentrations of 2 mg/ml desiccated Mycobacterium tuberculosis (Difco 231141) (to make complete Freund’s adjuvant, CFA) and 0.5 mg/ml myelin oligodendrocyte (MOG) peptide 35-55 (Genscript RP10245). Three injections were administered s.c. in the back skin (100 μl per site) such that each mouse received 150 μg total MOG. At the time of MOG immunization and 1 d later, 200 ng pertussis toxin (List Biological 181) was injected i.p. …

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

EAE induction in mice

… Eight-ten week-old C57BL/6 N female mice (Charles River Laboratories, Montreal, QC, Canada), housed under specific pathogen free conditions, 4/cage, on a 12:12 h light cycle were used in this study. EAE was induced by subcutaneous immunization with 100 μg myelin oligodendrocyte glycoprotein (MOG35–55) in emulsion of a 1:1 volume with complete Freund’s adjuvant (CFA, containing 4 mg/ml of heat killed Mycobacterium tuberculosis, Difco Laboratories, Detroit, MI, USA). The mice were also injected with intravenous Bordetella pertussis toxin (200 ng; PTX, List Biological Laboratories Inc., Campbell, California, USA) in 1X Phosphate Buffered Saline (PBS) at the time of, and two days 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)

Detection of PA and Anti-PA IgG in Serum

PA was measured in serum using PA-ECL screening assay kit MesoScale Discovery (L15CA-1), which utilizes the anti-PA antibody. To quantitate the levels of PA in each serum sample, a standard curve (0–100 ng/mL) was analyzed in parallel on each assay day by using recombinant PA (List Biological Laboratories, Inc., 171E). Test samples were assayed in duplicate.

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

2.1. RR-EAE Induction

SJL/JCrl female mice were purchased from Charles River (Calco, Italy). All mice were housed in pathogen-free conditions with access to food and water ad libitum. RR-EAE was induced in female wild type mice (7 weeks of age, weighing 18.5 ± 1.5 g) via a subcutaneous injection at two different sites in the right and left flanks with an emulsion (200 μL total) containing 200 μg of proteolipid protein peptides spanning amino acids 139–151 (PLP) (Espikem, Florence, Italy) in incomplete Freund’s adjuvant (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) supplemented with 600 μg Mycobacterium tuberculosis (strain H37RA; Difco). Mice were injected in the tail vein with 400 ng pertussis toxin (List Biological, Campbell, CA, USA) in 100 μL of phosphate buffer saline solution (PBS, pH 7.6) immediately and 48 h after the 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)

EAE induction

For active EAE induction, age- and sex-matched mice were immunized s.c. with MOG35-55 (Genscript, purity >98%) peptide (300 μg) emulsified in CFA containing 4 mg/ml heat-killed MTB (Cat#7001,Chondrex). 200 ng Pertussis toxin (Cat#181239A1, List Biological Laboratories) dissolved in PBS was administered i.p. on days 0 and 2. …

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

EAON Model

EAON was induced in an experimental autoimmune encephalomyelitis (EAE) mouse model (WT, n = 12; limitrin KO, n = 18), as previously described.4,5,23,24 Areas draining into axillary and inguinal lymph nodes of 7- to 8-week-old mice were subcutaneously injected with 200 µg myelin oligodendrocyte glycoprotein (MOG35–55 peptide fragment MEVGWYRSPFSRVVHLYRNGK; GL Biochem Ltd., Shanghai, China) emulsified in 100 µL solution containing 50% complete Freund’s adjuvant and 10 mg/mL heat-killed H37Ra strain of Mycobacterium tuberculosis (Difco Laboratories, Detroit, MI). Pertussis toxin (List Biological Laboratories, Campbell, CA) dissolved in PBS (200 ng/mouse) was administered intraperitoneally on days of immunization and again 48 hours later. …

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)

In Vivo Optic Nerve Transection and NSC Transplant

… Three days before tissue collection, animals received intravitreal injections of cholera toxin subunit B (CTB; 2 µL, 1 µg/µL; List Biological Labs, Campbell, CA, USA) to anterogradely label RGC axons. Animals were perfused transcardially with cold PBS, followed by 4% PFA, and post fixed in 4% PFA overnight. The eyes were dissected and stored in PBS. The optic nerves were dissected and cryoprotected in 30% sucrose.

Retina and Optic Nerve Immunohistochemistry

… Primary antibodies to CTB (1:1000; List Biological Labs), …

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