Bacterial Toxin Research Citations

… FoxP3DTR mice to receive transferred Treg cells were treated with 50 mg/kg diphtheria toxin (DT) (List Labs, 150) starting from 5 days prior to tumor inoculation and for every 3 days until time of euthanization. Mice were monitored daily for tumor growth. Tumor volume was measured through digital calipers along the x, y, and z axes and calculated as (xyz)/2. Mice were to be euthanized if tumors exceeded size limit agreed by the Institutional Review Board (IRB), however no animal reached this volume during the duration of the experiment. …

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

EAE immunization
8-12 weeks-old male and female mice, minimum of 3 per gender and genotype, were subcutaneously injected at three sites with 200 µg of MOG peptide 35–55 emulsified in Complete Freund’s Adjuvant (CFA) containing 400 µg of Mycobacterium (Cat#: EK-0111, Hooke Laboratories). On day 0 (D0) and 1 day after (D1) immunization, mice were intraperitoneally injected with 200 ng of pertussis toxin (Cat#: 180, List Biological Laboratories). …

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

… Tetanus toxoid fluorophore conjugation Tetanus toxoid (191B; List Labs) was conjugated using the Lightning Link FITC conjugation kit (ab102884; Abcam) per manufacture’s instruction. …

• Product #191 – Tetanus Toxoid from Clostridium tetani

… Male 6-8 weeks old C57BL/6J/Sja mice (Hunan Slack King Experimental Animal Company) were maintained under specific pathogen-free conditions. Ultra Pure LPS (Escherichia coli 0111:B4, LIST Biological Laboratories, Campbell, USA) was dissolved in sterile normal saline and injected intraperitoneally (1 mg/kg) every two days to induce chronic endotoxemia for 4 weeks, followed by an injection every two weeks until the 12th week. …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

Transduction of mouse astroglial cells with bONE-GO sensor and BRET measurements

… Kinetic traces are represented as change in BRET after subtraction of the baseline signal measured for 30 s before GPCR stimulation [ΔBRET·103 (baseline)]. Where indicated in the figures or figure legends, cells expressing Gαi bONE-GO WT were treated overnight with 0.1 μg/ml pertussis toxin (List Biological Labs, cat#179A). For the concentration-response curve presented in Fig. 7C, the average [ΔBRET·103 (baseline)] of the “Buffer” condition was first subtracted from all traces, and then the area-under-curve (AUC) was calculated in Prism (GraphPad) for each trace, followed by curve fit to a 3-parameter sigmoidal equation.

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)

Immunohistochemistry

Mice were deeply anesthetized with a mixture of isoflurane and oxygen and received an i.p. lethal dose of exagon (300 mg/kg) and lidocaine (30 mg/kg). Mice were perfused transcardially with phosphate-buffered saline (PBS 1X) and incubated (48 h/4 °C) in 4% paraformaldehyde. Coronal slices were cut at 50 μm and washed three times in PBS 1X before incubation in the blocking solution containing 0.03% Triton X-100 and 10% donkey serum or goat serum. … or with a goat anti-CTb antibody (1/10000, list labs #703), or a rabbit anti-GFP primary antibody (1/1000; Millipore, AB3080), a mouse anti-GAD67 primary antibody (1/500; Millipore MAB5406), a guinea pig anti-PV primary antibody (1/1000, synaptic system, cat#195004). …

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

Active EAE Induction and immunization to assess the myelin-specific recall T cell response

Mice were injected subcutaneously on the left and right sides of the chest with 50 ml of emulsion/side containing either 100 mg
MOG35-55 or PLP139-151 (Genemed Synthesis) in complete Freund’s adjuvant (CFA) that contained pestle-ground 2 mg/ml heatkilled Mycobacterium tuberculosis H37Ra in incomplete Freund’s adjuvant (Difco Laboratories). For EAE induction in C57BL/6J
mice, mice were provided 100-200 ng Bordetella pertussis toxin (PTX) (List Biologicals) i.p. on day 0 and day 2 post-injection.
Each new lot of PTX was titrated in preliminary EAE studies prior for use in experiments. …

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 induction. Six- to 8-week-old Vegfb WT and cKO female mice were used in the EAE model. EAE was induced by subcutaneous immunization with MOG35–55 peptides (200 μg/mouse, catalog T510219, Sangon Biotech) emulsified in complete Freund’s adjuvant (catalog F5881, Sigma-Aldrich), followed by administration of pertussis toxin (200 ng/mouse, PTX, List Biological Laboratories Inc.) on days 0 and 2 as described above. …

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)

Mouse phrenic nerve hemidiaphragm assay

… Subsequent inhibition of muscle contraction was considered an indication that the contraction response was due to ACh released by nerve stimulation (data from preparations with inhibition below 95% were discounted). The preparations were then washed 3× for 5 min and after a further stabilization period of 20 min, 1 ml of 10x BoNT (final concentration 10 pM for BoNT/A and BoNT/B (both LIST Biological Laboratories, Campbell, USA), and recombinant (r)BoNT/X, and 100 pM for LHN/X, in KHB supplemented with 0.05% (w/v) gelatin type A (Sigma) was added to the tissue bath and electrical stimulation continued until muscle contraction was ablated. …

… Frozen stocks of LPS (List Biological #R595) or PAM3 (Invivogen #vac-pms) were thawed and sonicated for 5 min before dilution in BMDM medium. The diluted stimuli were again sonicated for 5 min before addition to BMDM cultures for a final concentration of 10 ng/mL for LPS and 300 ng/mL for PAM3. CIM cells were stimulated with 100 ng/mL LPS. Unless otherwise indicated, cells were incubated with 25HC (5 ?M) (Sigma #H1015), OSW-1 (0.1 nM) (Cayman Chemicals #30310), …

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