Bacterial Toxin Research Citations

Primary Human Monocytes:

…Cells were allowed to adhere to the plates in an incubator at 37°C, 5% CO₂, and 95% relative humidity for 2 hrs. After 2 hrs, the cell culture medium was aspirated, cells were washed with fresh medium and incubated with 5 ng/mL of ultrapure LPS from Escherichia coli O111:B4 (List Biological Laboratories, Inc., Campbell, CA) over night for priming. …

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

Materials:

… Lipopolysaccharides from Salmonella minnesota R595 (Re) were purchased from List Biological Laboratories Inc. (Campbell, CA).  Biotinylation of S. minnesota R595 (Re) lipopolysaccharides was performed using EZ-Link Sulfo-NHS-LC-Biotin…

Binding Assay to Lipopolysaccharide-immobilized Sepharose:

Sepharose coupled to streptavidin was incubated for 2 h at 4 C in the presence or absence of 10 g of biotinylated lipopolysaccharides in 1.0 ml of TBS1, and Sepharose, obtained by centrifugation at 6,000 g for 5 min was washed with TBS1. …

Amidase Activity of Recombinant Proteins Activated by Lipopolysaccharides:

Recombinant proteins were incubated in the presence of various concentrations of lipopolysaccharides in 100 mm Tris-HCl, pH 8.0, at 37 C for 20 min. …

Lipopolysaccharide-binding Assay with a Surface Plasmon Sensor:

Biotinylated lipopolysaccharides (2.3 m in 10 mm Hepes-NaOH, pH 7.4, containing 150 mm NaCl) were immobilized on a sensor chip SA of a BIAcore X system…

• Product #304 – LPS from Salmonella minnesota R595 (Re)

… For regulatory T cell depletion, wild-type control or Foxp3-DTR mice were injected intraperitoneally with 1.5 g of DT (Calbiochem, San Diego, CA, USA or List Biological Laboratory, Campbell, CA, USA) on days 0, 1, 4, and 7 and examined on day 8. CD4+, CD25+, Foxp3+ …

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

(Transwell Migration Assays) – Pertussis toxin (PTX) was purchased from List Biological Laboratories, Inc. (Campbell, CA, USA). …

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:

Escherichia coli O111 B4-derived LPS was obtained from List Biological Laboratories, Inc. (Campbell, CA, USA).

Northern blot analysis and Cox2 mRNA inhibition:

The procedure employed was similar to that reported previously (14). Briefly, 106 cells were placed in Falcon 5-cm-diameter dishes (Becton Dickinson Labware, Franklin Lakes, NJ, USA) and pre-treated for 30 min with each of the phenol-related compounds at a concentration of 106 M, 105 M or 104 M. They were then incubated in the presence or absence of LPS (100 ng/ml), and their total RNA was prepared 3 h later by the acid guanidine-phenol-chloroform procedure (15). …  Statistical analyses were performed using Students t-test. Inhibition of LPS-stimulated Cox2mRNA expression byp-cresol, p-cresol dimer, pHA, pHA dimer and BHA in RAW264.7 cells was carried out as follows: The cells were pretreated for 30 min with or without addition of 105Mp-cresol, pHA,p-cresol dimer, pHA dimer or BHA, and then incubated with or without LPS at 100 ng/ml for 3 h. Total RNA was prepared and Cox2 mRNA expression was confirmed by northern blot analysis.

Preparation of nuclear extract and microwell colorimetric Nfb assay:

Nuclei were extracted and prepared for the microwell colorimetric Nfb assay. In brief, the cells in 5-cm-diameter Falcon dishes (106cells per dish) were pretreated for 30 min with or without the indicated doses of the compounds, and then treated with LPS at 100 ng/ml for 1 h. …Inhibition of LPS-stimulated Nf-b activation byp-cresol,p-cresol dimer, pHA, pHA dimer or BHA in RAW264.7 cells was carried out as follows: The cells were pretreated for 30 min with or without addition of 105 M p-cresol, pHA, p-cresol dimer, pHA dimer or BHA, and then incubated with or without the LPS at 100 ng/ml for 1 h. Nuclear extracts were prepared and used in a TransAM (Active Motif) ELISA-like assay to quantify the Nfb p50, p52, p65 and RelB DNA-binding activity. …

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

… To determine whether antisera induced by CTB or CTBCOMP fusion protein with or without heat treatment inhibit CT from binding its receptor GM1, 100 µl of the serially diluted antisera were incubated with 100 µl of CT (0.4 µg/ml in PBS, List Biological Laboratories, Inc …

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

Murine experimental autoimmune encephalomyelitis (EAE):

…Active induction of EAE was performed with a subcutaneous injection of each mouse with 200µg of myelin oligodendrocyte glycoprotein (MOG) 3555 peptide… Two intra-peritoneal (i.p.) injections of Bordetella pertussis toxin (PTX; List Biological, Campbell, CA) at 200 ng pe mouse was given at the time of immunization…

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)

Inhibition of Rho GTPase activity:

To study cellular phenotypes independent of GTPase activation, cells were treated with either Clostridium difficile toxin B (TcdB) or C3 transferase to irreversibly inactivate either RhoA, Rac and Cdc42 or RhoA, respectively. Cells were treated wither with 200 ng/ml TcdB (List Biologicals) or 1 g/ml cell-permeable C3 (Cytoskeleton) for 4 hours. Attachment experiments were carried out immediately after toxin treatment.

• Product #155 – Toxin B from Clostridium difficile

Chemicals:

… cholera toxin was from List Biological Laboratories, Inc. (Campbell, California, USA) …

Mouse models of cholera and intestinal fluid absorption assays

To investigate the in vivo effect of diclofenac on CT– and V. cholerae-induced intestinal fluid secretion, mice (30–35 g, ICR strain; The National Laboratory Animal Center, Salaya, Nakornpathom, Thailand) were fasted for 24 h before experiments. After anesthesia by an intraperitoneal injection of thiopenthal sodium (50 mg/kg), an abdominal incision was made and 3 closed ileal loops (2–3 cm in length) were made by ligations and then instilled with 100 µl of phosphate-buffered saline (PBS) or PBS containing CT (1 µg) or V. cholerae (classical O1 569B strain of V. cholerae at 107 CFU/loop). This strain of V. cholerae was used since it has been known to produce large amounts of CT and cause consistent intestinal fluid secretion in adult mouse closed-loop models [17]. Body temperature of mice was maintained at 36–37°C for the entire period of operation using heating pads. After abdominal closure by sutures, mice were intraperitoneally administered with DMSO (control) or diclofenac (30 mg/kg), and allowed to recover from anesthesia. Four hours (for experiments using CT) or 12 hours (for experiments using V. cholerae) later, mice were anesthetized again, abdomen was opened and ileal loops were removed for measurements of intestinal fluid secretion from loop weight/length ratios. Then, mice were euthanized with an injection of thiopenthal sodium (150 mg/kg). …

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

… For SL administration, the HA1 protein was mixed with wild-type cholera toxin (wtCT) (List Biological Labo- ratories, Inc., USA) as adjuvant. …

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