Citations

Recording solutions

… PTX (List Biological Laboratories, Inc., 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)

… Pertussis toxin (List Biological Lab, USA) (300 ng) solved in 100 µl PBS and was administered intraperitoneally,   2 h after immunization. The pertussis toxin injection was repeated the next day. …

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)

Induction and treatment of EAE model:

… Age matched mice were subcutaneously immunized with peptide MOG 35-55 (GL Biochem, Shanghai, China) mixed 1:1 with complete Freund’s adjuvant (Sigma, St. Louis, MO, USA), followed by pertussis toxin (200 ng; List Biological Laboratories, Campbell, CA, USA) administration i.v. on day 0 and day 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)

Llama immunization, serum fractionation and serology:

Recombinant TcdB (aa 17512366; hereafter referred to as TcdB_1751-2366) was a generous gift from Dr. Kenneth Ng (University of Calgary, Calgary, AB, Canada). A llama was immunized with 100 g of TcdB_1751-2366 per injection, following a similar immunization schedule and adjuvanted as previously reported [29]. Serum from blood drawn 42 days post immunization was tested for binding to TcdB_1751-2366 by ELISA essentially as described [10]. Serum was fractionated according to established protocols with protein G and protein A affinity columns [30] and conventional and heavy-chain IgG fractions tested for TcdB_1751-2366 binding by ELISA [10]. The ability of polyclonal fractions to neutralize TcdB VPI 10463 (List Biological Laboratories) was examined by Vero cell toxin inhibition assays, essentially as described for TcdA [19] with minor modifications. Vero cell monolayers were incubated with a final TcdB concentration of 10 pM (2.7 ng/mL) or 30 pM (8.1 pg/mL) and 1 M of day 42 fractionated serum for 72 h at 37C and 5% CO₂ before addition of WST-1 cytotoxicity reagent (Roche, Mississauga, ON, Canada) for 30 min and subsequent absorbance measurement at 450 nm.

• Product #155 – Toxin B from Clostridium difficile

… The sections were sequentially incubated for 3 days at 4C with goat anti-CTB antibody (1:10,000 dilution; List Biological Laboratories, Campbell, CA, USA, Cat# 703 subunit B from Vibrio cholerae type Inaba 569B, which was obtained from List Biological …

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

List Labs has provided Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae) and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt).  The former has been discontinued.  The latter may be substituted for the former, if the researcher takes into account the difference in the buffer in which each product is provided and the amount of protein per vial.  When Product #103B was reconstituted with 200 l of water, it contained 1 mg of protein in 0.05 M Tris, 0.2 M NaCl, 0.001 M Na₂EDTA, 0.003 M NaN_3, pH 7.5.  When Product #104 is reconstituted with 0.25 ml water, it contains 0.5 mg of Cholera Toxin B Subunit (choleragenoid) in 0.01 M Sodium Phosphate at pH 7.5.

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

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