Bacterial Toxin Research Citations

Cell surface binding assay

Binding of WT and mutated TcdB to cells was analyzed by the cell surface binding assay21. Briefly, cells were exposed to TcdB (10 nM) or TcdB mutants (10 nM) for 10 min at room temperature. Cells were washed three times with PBS and lysed with RIPA buffer (50 mM Tris, 1% NP40, 150 mM NaCl, 0.5% sodium deoxycholate, 0.1% SDS, with a protease inhibitor cocktail (Sigma-Aldrich). Cell lysates were centrifuged and supernatants were subjected to western blotting using chicken polyclonal anti-TcdB IgY (List Labs, #754A, 1:2000) and goat anti-chicken IgY H&L (HRP) (Abcam, ab97135, 1:2000) antibodies to examine the binding of TcdB mutants. Chicken polyclonal anti-actin antibody (Aves Labs, ACT-1010, 1:2000) was used for negative control.

2.4.2 Oral tolerance induction, sensitization and challenge

… Two days after the last oral administration with the indicated pre-treatment, mice received five consecutive weekly oral sensitizations with 20 mg whey plus 10 µg cholera toxin (CT; List Biological Laboratories, Inc.) as adjuvant in 0.5 ml PBS for Group 2–7 to break oral tolerance for whey, and 10 µg cholera toxin in 0.5 ml PBS for the non-sensitized mice in Group 1. …

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

2.3 Effect of drug treatment on hormone-induced cilia length control

The following drugs were used in this study: pertussis toxin (PTX; List Biological Laboratories Inc.); …

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)

Diphtheria-toxin-induced depletion of TRMs.

Depletion of TRMs was induced by intranasal instillation of 15 ng per mouse of diptheria toxin (DT, List Biological Laboratories).

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

… Additionally, each site was injected 88 ip with 100 ng of pertussis toxin (List Biological Laboratories, 181) immediately after MOG35-55 89 injection and again 2 days after immunization. …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

… LPS was obtained from List Biological Laboratories, INC. (#423, California, USA). …

In vitro stimulation:

At the day of experiments the cell medium was replaced by pre-warmed 500 µL DMEM/antibiotics per well. Cells were stimulated with LPS (10ng/ml), TNF-α (1, 20, 100 ng/ml), IL-1β (1, 20, 100 ng/ml), galectin-3 (5, 50 or 100 ug/mL), or the combinations of LPS (10 ng/ml) or TNF-α (100 ng/mL) with galectin3 (5 ug/ml or 100 ug/ml) for 24 h. Cell culture media from each well was collected and stored at -80°C until analysis. Each experiment was repeated at three independent times using 6 animals/repeat. Vehicle and treatment wells were situated on the same plate and two samples for each treatment were analyzed for every repeat. …

• Product #423 – ULTRA PURE LPS from Escherichia coli O55:B5

EAE induction

Active EAE was induced as described [17, 21]. Subcutaneous flank injection of 300µg MOG35 − 55 peptide (MEVGWYRSPFSRVVHLYRNGK, Millipore Sigma) in complete Freund’s adjuvant (CFA, Difco) containing 300µg Mycobacterium tuberculosis was performed on day 0 (D0), and supplemented by intraperitoneal injections of 500ng pertussis toxin (PTX, List Biological) on D0 and D2 to heighten the auto-immune reaction to MOG peptide [22, 23]. …

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 AND METHODS

A. Proteins and reagents

Anthrax Protective Antigen (PA, recombinant from B. anthracis) was purchased from List Biological Laboratories Inc. (Campbell, California, USA). …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Induction of experimental autoimmune encephalomyelitis (EAE)

Mice were immunized with 200 µg of synthesized MOG35–55 peptide (Anygen, Korea) in CFA (BD Difco) emulsion containing 400 µg of Mycobacterium tuberculosis (BD Difco) on day 0. Mixture or MOG35–55 peptide, CFA, and M. tuberculosis was subcutaneously injected into both right and left flank side of the mice. Mice were administered 400 ng of pertussis toxin (List Biological Laboratories Inc.) through intraperitoneal route 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)

Ablation of AgRP neurons

To ablate AgRP neurons, mice carrying the AgrpDTR/+ allele, were bilaterally microinjected with DT (0.4 ng/side/mouse, List Biological Laboratories, Campbell, CA) using a needle (Hamilton Small Hub RN 33 G, Reno, NV) connected with a 10 μl syringe (Hamilton 700 Microliter, Reno, NV), at a rate of 0.1 μl/min. A total 0.4 μl volume/side was delivered into the dlDRN2. To describe the expression profile of AgRPARC-dlDRN neurons, the vehicle (0.9% NaCl) or DT were injected into the dlDRN with AgrpDTR/+::NpyGFP mice. …

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