Bacterial Toxin Research Citations

Injection site visualization:

… In the other cases (S4-5, S11-13 and S17-18), the sections were first rinsed with PB 0.1M and then permeated with PB 0.1 M and 0.3 % Triton X-100. They were then incubated overnight in a choleragenoid antibody raised in goat (goat anti-CTb, lot no. 703, List Biological Laboratories, USA) diluted 1: 2000 in a blocking solution (PB 0.1 M, 5 % BSA). …

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

… Microinjection of CTB. Under general anaesthesia with isoflurane, a total of 3 µL 1% CTB (List Biological Labs, Campbell, California, USA) was used for injections at GB14, ST2 and ST6 in different rats (n=3 for each acupuncture point) …

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

EXPERIMENTAL PROCEDURES

Antigens and antibodies:

PTx (holotoxin) in glycerol was purchased from List Biological Laboratories, Inc. (Campbell, CA) …

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

… Cholera toxin was purchased from List Biological Laboratories. …

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

Cell stimulation:

RA synovial fibroblasts were seeded into 24 well plates and incubated at 37C, 5% CO₂ until confluent. The culture medium was replaced with 500l fresh complete RPMI 1640 medium (10% FCS, 400M L-glutamine, 2,000U/ml penicillin, 2mg/ml streptomycin, 1% v/v NEAA, 1% v/v sodium pyruvate, all from Sigma Aldrich, Gillingham, UK) prior to treatment with 10ng/ml of each of the cytokines IL-1, IL-17, IFN, TNF, TGF, IL-4 (all R&D Systems, Abingdon, UK), or IL-6 (Peprotech, New Jersey, USA) (20ng/ml), or TLR agonists 300ng/ml bacterial lipoprotein (bLP; InvivoGen, San Diego, US), 10g/ml poly(I:C) (InvivoGen, San Diego, US), or 100ng/ml LPS (List Biological Laboratories, Campbell, USA). The cells were incubated for 24hours at 37C, 5% CO₂ after which RNA was extracted for Gal9 expression as determined by Real time PCR.

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/

Induction of EAE and Tissue Collection:

EAE was induced in 810-week old WT or Nlrp12^/ female mice as previously described [19]. … Mice were also injected intraperitoneally on days 0 and 2 with 200 ng Pertussis toxin (List Biological Laboratories Inc., Campbell, CA, USA). After 3 weeks of immunization, mice were sacrificed, perfused with ice-cold phosphate-buffered saline (PBS) (Wisent, St. Bruno, QC, Canada) and the CNS tissues were collected.

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:

…Mice were subcutaneously immunized with MOG/CFA into two sites of dorsal flank followed by an intraperitoneal injection with pertussis toxin (400 ng; List Biological Labs, Inc., CA, USA, #180) at day 0 (immediately after immunization) and day 2 (48 hours later) …

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

Induction of EAE and peptide treatment:

EAE was induced in mice (8–12 weeks of age) as previously described21. Briefly, mice were injected subcutaneously with 200 µL of recombinant MOG35–55 (Biomatik Corporation), emulsified in incomplete Freund’s adjuvant (Sigma, Oakville, Canada) and supplemented with 4 mg/mL Mycobacterium tuberculosis (strain H37Ra, BD Biosciences, Mississauga, Canada) at four different sites with 50 µL per site. 200 ng of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA) was injected intraperitoneally (i.p.) on day 0 and 2 after 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)

Flow cytometry:

CD169 expression on MDMs was quantified by staining with Alexa647-conjugated mouse anti-human CD169 antibody (BioLegend, # 346006, 1:50) and analyzed with BD LSRII (BD). Geo mean fluorescence intensity (MFI) was calculated and normalized to that of uninfected (mock) MDMs. In some experiments, intracellular p24 was stained as described⁴⁹ using FITC- or RD1-conjugated mouse anti-p24 monoclonal antibody (KC57, Coulter, # 6604665 or # 6604667, 1:25). For T-cell exhaustion assay, MDMPBMC co-culture was harvested, stained with Zombie-NIR (BioLegend, #423105, 1:250) and a panel of makers designed for inhibitory receptor expression profiling⁵⁷, and analyzed with FACS ARIA (BD). For intracellular IFN- staining, MDMPBMC co-culture was unstimulated or stimulated with SEB (List Biological Laboratories) for 1h, followed by culture with brefeldin A (BioLegend) for another 5h, stained for inhibitory receptors as described above, fixed with FluoroFix Buffer (BioLegend), and stored in a staining buffer (2% NCS in PBS). Fixed cells were permeabilized with intracellular staining permeabilization wash buffer (BioLegend), stained with anti-human IFN- PE (BioLegend, # 506506, 1:200), and analyzed with FACS ARIA (BD). Data was analyzed with FlowJo software (FlowJo).

EAE:

… Pertussis toxin (List Biological Labs) was injected intraperitoneally (ip) on day 0 and day 2 of the immunization [6].  Mice were monitored each day for the development of clinical symptoms. …

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)