Bacterial Toxin Research Citations

Experimental procedures

Reagents:

… E. coli 0111:B4 LPS from List Biological (Campbell, CA); …

Murine in vivo exposures:

C57BL/6J female mice, 8-10 weeks old and weighing 18-22g, were used and were from Jackson Laboratories (Bar Harbor, ME). All experiments were performed in accordance with the Animal Welfare Act and the U.S. Public Health Service Policy on Humane Care and Use of Laboratory Animals after review and approval by the Animal Care and Use Committee of the NIEHS. Lentiviral shRNA (scramble control or Lrch4 specific; 6×107 TUs in 50 l saline) were delivered to the lung by oropharyngeal aspiration during isoflurane anesthesia, similar to past reports (31,32). Exposure to aerosolized LPS (300 g/ml, 30 min) was as previously described (33).

• Product #201 – LPS from Escherichia coli O111:B4

2.5. Immunization of mice and parasite challenge

Female BALB/c mice (five animals per group) were immunized by TCI by directly placing the antigens onto their shaved back [6]. The antigen doses determined separately in a dose ranging experiment were 50 μg of SLA or rLACK mixed with 5 μg of CT (List Biological Laboratories) adjuvant in a total volume of 50 μl. Control groups received only CT or PBS. …

2.6. Antibody measurement by enzyme-linked immunosorbent assay (ELISA)

Serum samples collected from immunized mice were analyzed by ELISA for the presence of IgG antibodies specific to rLACK and SLA, as previously described [27]. Individual samples were analyzed in triplicate at each time point and results plotted as an absorbance at 450 nm (OD at 450 nm) for SLA and rLACK. The CT titer was determined as previously described with some modifications [28]. Briefly, plates were coated with CT, washed and blocked in phosphate buffered saline (PBS) containing 0.5% gelatin (Sigma Aldrich) for 1 h at 37°C. …

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

… Delivery of LPS (Escherichia coli O111: B4, List Biological Labs, Inc) was performed using the tongue-pull maneuver in isoflurane- anesthetized mice, as described previously (18, 19). …

• Product #201 – LPS from Escherichia coli O111:B4

Monitoring Corneal Wound Resolution in Organ Culture:

Confetti mice were wounded as described above. The animals were euthanized and their eyes immediately enucleated. Paired (wounded and intact) globes were embedded in sterile 1.5% agarose and placed within the microscope’s imaging/incubation chamber, set to 37C and 5% CO₂ prior to immersion in defined keratinocyte serum-free medium (Gibco, Fremont, CA) containing 50 nM recombinant human epidermal growth factor (Peprotech, Rocky Hill, NJ), 500 nM cholera toxin (List Biological Labs, Campbell, CA) and 100 U/mL penicillin-streptomycin (Gibco). Eyes were imaged every 2 hr over 48 hr by light-sheet microscopy (Zeiss Lightsheet Z.1) using a 5/0.16 detection lens and 5/0.1 illumination lens as described above.

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

… Mice (n=6 per group) were intragastrically (ig) sensitized with 6 mg PE and 15 µg Cholera Toxin (CT, List Biological Laboratories, …

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

Bioreagents:

… Polyclonal anti-botulinum neurotoxin type A [#730A, -BoNT-A pAb (chicken IgY)], monoclonal anti-botulinum neurotoxin type A [#731A, -BoNT-A mAb (mouse IgG)], botulinum neurotoxin type A toxoid [#133L, BoNT-A], and recombinant botulinum neurotoxin type B light chain [#620A, BoNT-B were obtained from List Biological Laboratories. All lyophilized bioreagents were reconstituted and stored per manufacturer recommendations.

• Product #730A – Anti-Botulinum Neurotoxin, Type A (Chicken lgY)
• Product #731L – Anti-Botulinum Neurotoxin, Type A (Mouse lgG monoclonal F1-40), Frozen Liquid
• Product #133L – Botulinum Neurotoxin Type A Toxoid from Clostridium botulinum
• Product #620A – Botulinum Neurotoxin Type B Light Chain, Recombinant

Botulinum toxins (BoNTs) and general reagents:

…  Serotypes A, B, and E were also purchased from List Biological Laboratories as complex toxins and BSA was added during the reconstitution step of the lyophilized powder supplied, as recommended by the manufacturer. …

Table1

Table showing primary and secondary antibodies:

… Goat anti-CtB, List Laboratories 703, …

Immunouorescencestaining:

Tissues were obtained from a previous study [1].  We evaluated with immunolabeling (Table1) whether neurotrophin-3 treatment regulates Sema4C expression specically in TrkC+ afferent neurons, which include proprioceptive afferents. …

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

Drugs and reagents:

… Pertussis toxin (PTX) was from List Biological Laboratories, Inc. (Campbell, CA, USA). …

Dynamic mass redistribution assay:

Antagonist properties of ligands were measured by assessing the concentration response curve to NPS in the absence and in presence of a fixed concentration of compound. FR900359 was added 60 minute before NPS, rolipram was incubated for 90 minute before NPS, while PTX was added 24 hours before NPS. Maximum picometer (pm) modifications (peak measured at 60 minute time point) were used to determine agonist response after baseline normalization.

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)

Materials:

Cholera toxin was purchased from List Biological Laboratory, Inc, California, USA; …

Induction of fluid accumulation by cholera toxin (CT):

Fluid accumulation in mouse intestinal loops in response to CT was performed in ARH1^+/+ and ARH1^-/- mice [28, 39]. After mice were anesthetized, intestine was exteriorized through a midline incision. Two or three intestinal segments of about 4 cm length each were generated by ligation with nylon suture, and 0.2 ml of PBS or PBS containing either 0.5 µg CT or 5 mM 8-Bromo-cAMP were injected into each loop [28]. …

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