Bacterial Toxin Research Citations

… Cholera toxin was acquired from List biological labs (Campbell, CA, USA). To analyze cell death after drug treatment, gelatin, neural buffered formalin (NBF), triton X-100, bovine serum albumin (BSA) and 4’,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma Aldrich (St. Louis, MO, USA), while cell tracker deep red and CD324 …

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

STAR★Methods
Key resources table

Goat anti-Cholera Toxin B Subunit List Biological Laboratories

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

Active EAE
C57BL/6J mice were purchased from The Jackson Laboratory. The animals were housed in the animal facility at the Johns Hopkins University School of Medicine 12 h/12 h light/dark cycles and fed with standard food and water. Nlrx1−/− mice (C57BL/6J) were kindly provided by Dr. Denis Gris (Sherbrooke, QC, Canada) [20, 22]. We induced EAE in 9-week-old C57BL/6J WT and Nlrx1−/− mice with myelin oligodendrocyte glycoprotein (MOG35–55) immunization as previously described [27]. Briefly, mice were immunized on day 0 subcutaneously at two sites over the lateral abdomen with an emulsion of MOG35 − 55 (200 µg, JHU Peptide Synthesis Core) in complete Freund’s adjuvant (CFA) containing 400 µg of Mycobacterium tuberculosis H37RA (Difco Laboratories). Pertussis toxin (250 ng; List Biological Labs, Campbell, CA, USA) was intraperitoneally injected on days 0 and 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)

Treg cell transfer, diphtheria toxin, BrdU, and anti-amphiregulin administration

Thymi and spleen from 6–8-week-old female Foxp3RFP or Foxp3RFPRag2GFP were harvested and strained through a sterile 40 μm filter. Next, red blood cells were lysed and stained before sorting on a 70micron nozzle. 100.000 RFP+ thymic or splenic Treg cells or Rag2GFP+ or Rag2GFP- Thymic Treg cells were then injected retro-orbitally into mice 3 hours after SL-TBI. Diptheria toxin (DT; List Biological Laboratories) was administered at a dose of 50 ng/g in PBS via IP injections on 2 consecutive days. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Mouse Treatment

For depletion of macrophages, 1 day before tumor inoculation, mice were injected via the tail vein with 200 µL mouse−1 clodronate liposomes or control PBS liposomes (40337ES10 or 40338ES10, Yeasen Biotechnology) every 4 days. For depletion of CD8+ T cells, mice were injected intraperitoneally with anti-mouse CD8α antibody (12.5 mg kg−1, BE0061, BioXcell) or isotype antibody (BE0090, BioXcell) every 3 days after tumor inoculation. For depletion of Tregs, 2 days after tumor implantation, C57BL/6J Foxp3DTR mice were performed by intraperitoneal injection of diphtheria toxin (12.5 µg kg−1, 15047A1, List Labs) or PBS every 3 days. For neutralization of CCL20 in vivo, tumor-bearing mice were injected intraperitoneally with 10µg dose−1 anti-mouse CCL20 antibody (AF760, R&D Systems) or isotype every 3 days from the third day. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Induction and treatment of EAU.
Mice were injected subcutaneously with 100 μL hIRBP1–20 (GPTHLFQPSLVLDMAKVLLD, 2 μg/mL; Sangon Biotech) emulsified in 100 μL incomplete freund’s adjuvant (Sigma, F5506), which contained the mycobacterium tuberculosis strain H37Ra (2.5 mg/mL; BD Difco). Additionally, 200 ng pertussis toxin (List Labs, #180) was administered intraperitoneally twice (on day 0 and day 2) to each mouse after immunization. Neutrophils were depleted by intraperitoneally injecting 10 μg/g of the neutrophil-specific antibody Ly6G (Biolegend, 127649) or 10 μg/g rat IgG2A isotype control antibody (Biolegend, 400565) as isotype control for every 2 days (39). NET degradation was performed by daily subcutaneous administration of 150 U/mouse DNase I (Roche, 04536282001) in 0.9% sodium chloride solution or 0.9% sodium chloride solution as vehicle every day. …

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

2.2 EAE

Female WT and Fbln2−/− mice (10–12 weeks old) were injected subcutaneously with 50 μg MOG35–55 peptide (synthesized by Protein and Nucleic acid Facility, Stanford University, Stanford, California, USA) emulsified in complete Freund’s adjuvant (CFA) (Thermo Fisher Scientific) supplemented with 10 mg/mL heat-inactivated Mycobacterium tuberculosis H37Ra (MilliporeSigma). A total of 100 μL emulsion was injected at 2 sites into hind flanks. On the day of immunization and 48 h later, Pertussis toxin (300 ng per 200 μL; List Biological Laboratories) was injected intraperitoneally. …

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)

2.3 Tracers and injection site definitions

We used the iontophoresis method for PHAL and CTB co-injections, as described previously (Negishi et al., 2024). Glass micropipettes (tip diamaters: 12–20 µm) were filled with a cocktail containing 2.5% PHAL (Vector Laboratories, Inc., Newark, California, USA; catalog #L-1110) and 0.25% CTB (List Biological Labs, Inc., Campbell, California, USA; catalog #104) dissolved in 10 mM sodium phosphate solution. …

2.5 Immunohistochemistry

… We then incubated tissues in primary antiserum containing antibodies raised against PHAL (host: rabbit; dilution: 1:4,000; Vector Labs, Cat # AS-2224; RRID: AB_2313686) or CTB (host: goat; dilution: 1:10,000; List Biological Cat # 703; RRID: AB_10013220) in blocking solution for 48 h in 4°C. …

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

Tissue processing

Fixed tissue was cryoprotected with 30% sucrose in PBS and then cut into 20 µm-thick horizontal sections that were blocked in phosphate buffer (PB) containing 5% Donkey serum and 0.2% Triton X-100 for 60 minutes at room temperature. Tissue sections were incubated overnight at room temperature with primary antibodies in PBS and 5% Donkey serum and 0.2% Triton X-100. Sections were then incubated with appropriate Alexa Fluor (ThermoFisher) secondary antibodies (1:500 dilution) in PBS for 2 hours at room temperature. Sections were counterstained with DAPI and mounted with Fluoromount-G (ThermoFisher Scientific). The following primary antibodies and dilutions were used: mouse anti-NeuN (1:200, Millipore # MAB377)), rabbit anti-ATF-3 (1:200, Novus #NBP1-8581), mouse anti-laminin (1:50, Sigma-Aldrich #CC095-M), goat anti-CTB (1:200, List Biological Laboratories, #703), …

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