Bacterial Toxin Research Citations

2.2 Active experimental autoimmune encephalomyelitis induction and control immunization

aEAE was induced in both male and female mice by injection of myelin oligodendrocyte glycoprotein peptide 35–55 (MOG35-55 peptide, 200 μg per animal, Genscript) and complete Freund’s adjuvant (Incomplete Freund’s Adjuvant, Santa Cruz Biotechnology; supplemented with Mycobacterium tubercolosis, Difco). An emulsion of MOG35-55 and CFA was injected subcutaneously in mouse flanks and at the tail base at day 0; in addition, 300 ng of pertussis toxin (List Biological Laboratories, Campbell, CA) was injected intraperitoneally at day 0 and day 2. …

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)

TLR agonists were: … Monophosphoryl Lipid A (MPLA) (List Biologicals, Campbell, California), …

• Product #401 – LIPID A monophosphoryl from Salmonella minnesota R595

Pertussis Toxin List Biologic Cat# 180

Induction and evaluation of EAE

To induce EAE, eight- to 12-week-old mice were injected subcutaneously with an emulsion contains 200 μg MOG35-55 in complete Freund’s adjuvant (CFA) containing 500 μg of nonviable, desiccated Mycobacterium tuberculosis at a concentration of 10 mg/mL on upper and lower back. Mice were i.p. injected with 200 ng pertussis toxin (PT) 2 hours later, and then repeated 2 days after immunization. For adoptive transfer of EAE with myelin-reactive T cells, splenocytes were isolated from MOG TCR transgenic (2D2) or wild type EAE mice. After removal of red blood cells by ammonium-chloride-potassium (ACK) lysis buffer, splenocytes were collected and suspended in RPMI-1640 medium containing 10% FBS. T cells were stimulated with 10 μg/ml MOG35−55 peptide for 48 h, and then expanded in 5 ng/ml IL-2 and IL-7 for 5 days. …

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

… Pertussis Toxin from List Biological Laboratories was dissolved in water …

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)

Table 1Summary of primary antibodies and dilutions for the immunolabeling. IF: immunofluorescence. IHC: immunohistochemistry (peroxidase).

Antibody Host Antigen Manufacturer RRID Dilution
CTB Goat/Polyclonal Cholera Toxin B List Biological Lab., CA, USA AB_10013220 1:5000 (IF)

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

… All animals were kept on a 12h dark-light cycle and standard housing conditions with ad libitum food access. At postnatal day 5 (P5), C57Bl/6 mice were randomly assigned to 4 experimental groups: lipopolysaccharides (LPS) (1 mg/kg, #423, List Biological Laboratories, Campbell, CA, USA)-injected mice …

• Product #423 – ULTRA PURE LPS from Escherichia coli O55:B5

… For immunolabeling, sections were blocked with 10% horse serum (Thermo Fisher Scientific, Waltham, MA, USA) in 0.1 M TBS, followed by the overnight incubation with primary antibody against choline acetyltransferase (ChAT, goat, 1:200, AB144P; Sigma-Aldrich, St. Louis, MO, USA) or CTB (goat, 1:500, 703; List Biological Laboratories, Inc., Campbell, NJ, USA) at 4 °C. …

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

2.4. EAE Induction and Scoring

Eight-week-old mice were immunized subcutaneously on the back with two injections of 150 μg myelin oligodendrocyte glycoprotein peptide (MOG35–55, GenScript, Piscataway, NJ, USA) emulsified in complete Freund’s adjuvant (CFA; Difco Laboratories, Franklin Lakes, NJ, USA) containing 2.5 mg/mL heat-killed Mycobacterium tuberculosis. Mice were further injected intraperitoneally with 200 ng pertussis toxin (List Biological, Campbell, CA, USA) in 0.1 mL of PBS at 0 and 48 h postimmunization with MOG35–55. …

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 of EAE

EAE was induced with 50 μg of myelin oligodendrocyte glycoprotein fragment MEVGWYRSPFSRVVHLYRNGK (MOG35–55 peptide; #crb1000205n Cambridge Research Biochemicals) mixed with incomplete Freund’s Adjuvant (#263910, BD) containing 4 mg/ml Mycobacterium tuberculosis TB H37 Ra (#231141, BD), at a ratio of 1:1 (v/v). Mice received two subcutaneous injections of 100 μl each of the MOG/CFA mix. Mice received a single intraperitoneal injection of pertussis toxin (#180, List Biological Laboratories) at a concentration of 1 ng/μL in 200 μL of PBS. A second injection of pertussis toxin at the same concentration was inoculated two days after the EAE induction. …

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

… (CFA) at a dose of 100 µg per mouse, followed by the administration of pertussis toxin (PT) (200 ng per mouse through intraperitoneal injection (IP); List biological …

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)