Bacterial Toxin Research Citations

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… Pertussis toxin List Biological Laboratories 180 …

In vivo induction of EAE

Induction and assessment of EAE were performed as previously described with minor modifications (Coll et al, 2015; Shi et al, 2016). Our study exclusively examined female mice because the disease modeled is only relevant in females. Each hind flank of 8–10 weeks female C57BL/6J mice were injected subcutaneously with 250 μg of myelin oligodendrocyte glycoprotein (MOG) peptide 35 to 55 emulsified in complete Freund’s adjuvant containing (0.4 mg/mouse) heat-killed Mycobacterium tuberculosis (4 mg/ml) at two different sites, followed by an intraperitoneal injection of 500 ng of pertussis toxin. On day 2, mice were given a second injection of 500 ng of pertussis toxin. Mice were assigned randomly to two groups and respectively administered with THL (2.5 mg/kg) or vehicle control (DMSO/ PBS) via intraperitoneal every second day starting at the induction of the disease on day 0. …

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

MATERIALS AND METHODS
Pertussis toxin List Biological Laboratories Catalog #179A

AT1R Ligands and Inhibitors

Angiotensin II was purchased from Sigma-Aldrich and TRV120023 was synthesized by GenScript. Both ligands were reconstituted in ultrapure water, and aliquots were stored at −20°C. FR900359 was purchased from Cayman Chemical. Pertussis toxin was purchased from List Biological Laboratories.

• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

STAR★Methods
Key resources table

… Pertussis toxin List Biological Laboratories Cat#180 …

EAE induction
For EAE induction, mice were injected subcutaneously at two sites on the back with 200 mg MOG35–55 peptide (synthesized by Toray Research Center, Tokyo, Japan) and 1 mg heat-killed Mycobacterium tuberculosis H37RA emulsified in complete Freund’s adjuvant (CFA; BD Difco, KS, USA). Briefly, 400 ng of Pertussis toxin (PTX; List Biological Laboratories, CA, USA) was injected intraperitoneally on days 0 and 2 after immunization. Two control groups were also added (no PTX control with MOG35–55/CFA and no MOG35–55/CFA control with PTX). EAE was clinically scored daily (0, no clinical signs; 0.5, tail weakness; 1, partial tail paralysis; 1.5, severe tail paralysis; 2, flaccid tail; 2.5, flaccid tail and hindlimb weakness; 3, partial hindlimb paralysis; 4, total hindlimb paralysis; 5, hind and foreleg paralysis). The mice given a score of 5 were euthanized at this point.

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

… 300 ng pertussis toxin (PTX) (List Biological Laboratories, #180) was injected i.p. at days 0 and 2 after immunization. EAE animals were scored as previously described. …

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

… cholera toxins (List Biological Laboratories, USA) and 1% v/v A/A. All of the cells were …

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

Binding assays
A high-binding 96-well ELISA plate (Costar) was incubated overnight at 4°C with 0.2 µg/mL PT (List Labs) in PBS, pH 7.4 or nothing (no coat). The plate was blocked with PBS-T-milk (PBS, pH 7.4 with 0.05% Tween-20 and 5% non-fat dry milk) for 1 hr at 25°C. Antibodies diluted to 5 µg/mL in PBS-T-milk were added to both the PT-coated and uncoated wells of the plate in duplicate and serially diluted 5-fold seven times. …

CHO cell clustering assay
Enzymatically active PT (List Labs) was diluted to 10 pM in CHO-K1 culture medium (high glucose DMEM with 10% fetal bovine serum and 1× penicillin and streptomycin). This PT diluent was used to prepare dilutions of antibodies hu1B7, hu1B7 S57A/Q62A/Q65A, or human IgG1 isotype control. The human IgG1 isotype control was also prepared in CHO-K1 culture medium without pertussis toxin (Isotype Control – no PT). The highest concentration of each antibody was 200 nM, with seven additional 2-fold dilutions prepared in wells of a black-walled, clear bottom 96-well plate (Agilent). The PT and antibody mixtures were incubated at 37°C for 30 min, then an equal volume of CHO-K1 culture medium with 104 CHO-K1 cells per well were added to the plate, diluting the pertussis toxin and antibodies by 2-fold. …

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 Islet-Activating Protein List Biological Laboratories Cat#: 180 …

Experimental autoimmune encephalomyelitis model
Fourteen days after tamoxifen treatment, mice were immunized subcutaneously with 100 mL of emulsified Complete Freund adjuvant
(BD Difco) supplemented with 4 mg/mL Mycobaterium tuberculosis H37Ra (BD Difco) and 200 mg MOG35-55 peptide
(MEVGWYRSPFSRVVHLYRNGK, Genemed Synthesis) and received intraperitoneal injections of 200 ng Pertussis Toxin from
Bordetella pertussis (List biological Laboratories) at the time of immunization and 48h later. Clinical disease was assessed by the
scoring of ascending hindlimb paralysis as follows: no signs, score 0; paralysis of tail, score 1; hindlimb weakness, score 2; paralysis
of one hindlimb, score 3; paralysis of both hindlimb, score 4; and moribund mouse, score 5.

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

EAE induction
Active EAE was induced by injection of 200 μg of myelin oligodendrocyte glycoprotein epitope MOG35-55 (#RP10245, Genscript) emulsified in complete Freund’s adjuvant (CFA) (#231131, BD). Mice received two subcutaneous (s.c.) injections into the flank and intraperitoneal injection (i.p.) of 200 ng pertussis toxin (PT) (List Biological Laboratories, #179A) the same day and two days after. …

• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Experimental autoimmune encephalomyelitis (EAE)
Mice were immunized 12 weeks after tamoxifen administration at the age of 15–16 weeks. MOG35-55/CFA was prepared by mixing 1 mg/ml MOG35-55 (GenScript) in PBS with Complete Freund’s Adjuvants (CFA, BD Biosciences) containing mycobacterium tuberculosis H37RA (BD Biosciences). 50 μg of the MOG35-55/CFA emulsion were injected subcutaneously into the tail base. Mice were also injected intraperitoneally with 150 ng pertussis toxin (List Biological Laboratories) in PBS at the day of immunization and two days 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)

Measurement of C. difficile TcdB concentration and sporulation capacity
TcdB concentration was quantified as previously reported [Citation37]. All 176 C. difficile isolates were cultured in brain heart infusion (BHI) broth at 37 ℃ for 48 h in an anaerobic chamber. TcdB concentration in BHI was measured and calculated using a formula according to a panel of purified TcdB standards (List Biological Laboratories, Campbell, CA, USA) with known concentrations ranging from 0.02 to 200 ng/μL. …

• Product #155 – Toxin B from Clostridium difficile