Bacterial Toxin Research Citations

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

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)

EAE Experiments

EAE was induced in male S1P1 -GFP signaling mice and GFP reporter mice aged 8-12 weeks. According to methods defined by Tsai et al [30, 31], mice were immunized with an emulsion containing 200 micrograms of Complete Freund’s Adjuvant (CFA), 100 micrograms of myelin oligodendrocyte glycoprotein (MOG35-55) peptide35–55, and 200 nanograms of Bordetella pertussis toxin (lot# 181236A1, List Biological Laboratories) on days zero and two. The spleen tissues were collected after eight days of immunization which marked the onset of the clinical EAE symptoms.

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

… Additionally, each site was injected 88 ip with 100 ng of pertussis toxin (List Biological Laboratories, 181) immediately after MOG35-55 89 injection and again 2 days after immunization. …

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

Induction and Evaluation of EAE

… On days 0 and 2, immunized mice received an intraperitoneal injection of 200 ng pertussis toxin (PTX, Cat: 181, Lot: 181238A1, List Biological Labs, CA, USA). …

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

… Pertussis toxin (salt free; 250 ng; List Biological Laboratories, Inc.) was injected intraperitoneally on day 0 and day 2 post-immunization. …

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

2.8 Adoptive transfer

For passive EAE, mice were euthanized 10 days after MOG35–55 immunization, and spleen and lymph nodes were collected as previously described [27]. Single cells were incubated for 3 days in the presence of 25 µg/mL MOG35–55, 10 ng/mL recombinant murine IL-23 (R&D, 1887-ML-010), and 2 ng/mL IL-2 (R&D, 402-ML-020) at 1 × 107 cells/mL. CD4+ T cells were purified by CD4+ T cell isolation kit and 1 × 106 cells per mouse were transferred via intravenous injection. Pertussis toxin (200 ng/mouse, List Biological Laboratories, 181) was injected intraperitoneally (i.p.) on days 0 and 2. EAE score was assessed by daily recording using a 0–5 scale.

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

… Pertussis toxin List Biologicals Cat# 181 …

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

EAE induction and in vivo treatment

Immunization: EAE was induced in 8-10 week-old naïve C57/B6 or female SJL/J mice by subcutaneously injection (s.c.) over four sites in the flank with 200 µg MOG 35-55 or PLP 139-151 respectively (C S bio) in an emulsion with CFA (DF3113605 Thermo Fisher Scientific). 200 ng pertussis toxin (181 List Biological) per mouse in PBS was injected i.p. at the time of immunization and 48 h later for C57/B6 mice. Female SJL/J mice only received one injection of pertussis toxin at the time of immunization. …

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

… 200 ng Pertussis-Toxin (List Biological Laboratories, Hornby, Ontario, Kanada; Kat-Nr. #181) in 500 µl steriler phosphatgepufferter Salzlösung (phosphate-buffered saline = PBS) (Dulbecco´s PBS; Sigma-Aldrich) …

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