Bacterial Toxin Research Citations

Induction of EAU

Purified whole bovine IRBP (150 µg) combined with IRBP peptide1–20 (300 µg; Anaspec, Fremont, CA) was emulsified in complete Freund’s adjuvant (CFA) containing 2.5 mg/ml heat-killed Mycobacterium tuberculosis H37RA (Difco, Detroit MI) as reported60 and was injected sub-cutaneously in the morning (9–11 a.m.) of the day of immunization. Mice were also intraperitoneally (i.p.) injected with 0.5 µg Bordetella pertussis toxin (PTX, List Biological Laboratories, Campbell, CA). …

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)

5.2. Retrograde tracer injections

Rats were anaesthetized with an intraperitoneal injection of ketamine (40 mg/kg) and xylazine (2.5 mg/kg). The head was shaved and the rat was placed in a stereotaxic frame in the “flat skull” position. Following a subcutaneous injection of local anaesthetic, an incision was made along the midline of the scalp and using a dental drill a small craniotomy was made on the midline of the skull, 3 mm caudal to bregma. Under stereotaxic guidance, using Paxinos and Watson (2005) as a reference, a glass micropipette (tip diameter 5-10um) filled with Cholera-toxin subunit B (CTB, List Biological Laboratories) was lowered through a small hole in the dura, into the paraventricular thalamus to a depth of 6 mm below the exposed cortical surface. Over a 20-minute period, 50 nl of CTB was deposited via an automated air-pressure system (9 ms air pulses). …

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

Neuroanatomical experimental procedures

Following the 2 h cold or warm exposure, rats were deeply anesthetized with pentobarbital (80 mg/kg i.p.) and transcardially perfused with saline followed by 4% paraformaldehyde (PFA). The brains were removed and post-fixed in 4% PFA for 2 h and equilibrated overnight in 20% sucrose in 10 mM sodium phosphate buffered saline (PBS; pH 7.4) with 0.01% sodium azide. Serial coronal Sects. (30 µm) were cut with a freezing-stage microtome, collected sequentially in 6 sets, and stored in cryoprotectant with 0.01% sodium azide at − 20 °C. Sections containing the preoptic area were pre-incubated for 3 h in an antibody dilution solution (ADS) containing PBS, 0.3% Triton-X 100, 0.25% carrageenan, 0.02% NaN3, and 1% normal donkey serum, and incubated overnight at room temperature in ADS containing the primary antibodies for c-Fos (rabbit anti-c-Fos , 1:10 K, Calbiochem) and for CTb (goat anti-CTb, 1:10 K, List Biologicals). …

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

… following intraperitoneal injections of 200 ng of pertussis toxin (List Biological Laboratories, Campbell …

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)

2.2. Experimental Design—Sensitization and Challenges

Mice were sensitized and treated as described previously [32]. In short, after an acclimatization period of 5 days, mice in the sensitization group were sensitized intragastrically (i.g.) with 20 mg whey protein (DMV International, Veghel, The Netherlands) and 15 µg cholera toxin (List Biological Laboratories, Campbell, CA, USA), which was used as an adjuvant, in 0.5 mL phosphate-buffered saline (PBS), once a week, for 5 consecutive weeks.

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

… All animals received 200 ng pertussis toxin (List Biological, Campbell, CA, USA) in 0.1 ml PBS by intraperitoneal injection at 0 h and 48 h after immunization. Severity of EAE was scored …

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)

Stereotaxic surgery and neuronal labeling

For retrograde tracing, eight week old wild-type ICR mice were used. Mice were anesthetized with isoflurane and attached to the stereotaxic frame. Small hole was drilled into the skull and unilateral, intracranial injections of 300nl of 0.2% Choleratoxin B subunit (CtB; List Biological Lab.Inc. Cat#104) was injected at the speed of 50nl/min using a microinjector (UltraMicroPump III, World Precision Instruments) and microsyringe (Hamilton, Cat#7803-06). Stereotaxic coordinates were measured from bregma in mm. To trace the LDTg neurons, CtB was injected into VTA: −3.1 (AP); 0.36 (ML); −4.65 (DV). Four to six days after the injections mice were intracardially perfused with 4% PFA and brains were collected, thereafter brains were fixed in 4% PFA for 24 h. 100 μm vibratome sections were cut for IHC stainings. Sections were blocked for 1 h in 10% donkey serum in PBS containing 05% Triton X-100 (PBS-T), followed by incubation overnight at room temperature with following antibodies diluted in PBS-T: rabbit anti-Nkx6-1 (1:400, Novus Biologicals); goat anti-CtB (1:5000, List Biologicals #703); mouse anti-TH (1:1000, Merck Millipore) and DAPI. Secondary antibodies conjugated to Alexa 488, 594, or 647 were used for detection. ProLong Gold anti-fade mounting media was used for mounting (Thermo Fisher Scientific, P36930).

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

EAE induction

Active EAE was induced following previously published procedures [17, 18]. Briefly, 8–10-week-old female mice were injected subcutaneously with 100 μg of murine myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) (EZBiolab), in complete Freund’s adjuvant (CFA) with 4 mg/mL Mycobacterium tuberculosis (DIFCO Laboratories). Mice also received 400 ng of pertussis toxin (LIST Biological Laboratories) intraperitoneally at the time of immunization and 48 h 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)

2.2 In vivo mouse model of dWMI

… After a recovery period of 75 min pups were exposed to 35 min of hypoxia (6% O2) in a temperature‐controlled hypoxic incubator (35.8–36.0°C). Directly following hypoxia, pups were i.p. injected with 1 mg/kg LPS (List Biological Laboratories, Campbell, CA) dissolved in 0.9% NaCl. …

Author did not specify which List Labs LPS product was utilized in their research.
List Labs provides the following LPS products: https://listlabs.com/product-information/lipopolysaccharides/

… For each assay, positive controls were obtained by stimulation with staphylococcal enterotoxin B superantigen (1 μg/mL, List Biological Laboratories, Inc, Campbell, CA). …