Bacterial Toxin Research Citations

Diphtheria toxin (DT) and puromycin treatments in vitro

Transfected HEK293 cells for human diphtheria toxin receptor (hDTR) mutagenesis were selected with 20 ng/ml DT (List Labs, #150) from day 3 after electro-transfections. Cell clones mutated in hDTR-sgRNA-1 or hDTR-sgRNA-2 targeted regions were cultured with DT-supplemented growth medium from day 1 after cell sorting (day 0 refers to the day of cell sorting). Transfected PFFs were selected with 300 ng/ml puromycin-containing (MPBIO, 219453925) medium for 48 h after electro-transfections of 12 h. Both DT and puromycin-supplemented growth medium were exchanged daily until negative control cells died completely.

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

EAE induction.

EAE was induced by subcutaneous immunization of mice into the flanks with an emulsion of MOG35-55 peptide (150 μg) emulsified in complete Freund adjuvant supplemented with 4mg/ml of M. tuberculosis extract H37Ra (Difco). In addition, the animals received 200 ng of pertussis toxin (List Biological Laboratories) i.p. 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)

Whey Sensitization/Challenge Protocol and Assessment of Anaphylaxis

The model of cow’s mike allergy-induced autism was established as described by de Theije’s et al, with modifications.37 Whey is a milk protein fraction composed of several milk allergens (β-lactoglobulin, α-lactalbumin, bovine lactoferrin, bovine serum albumin, and bovine immunoglobulins), some of which are major allergens, and used to induce our food allergy model.38 After 1 week acclimation, mice were sensitized intragastrically (i.g) with 20 mg whey (Hefei Bomei Biotechnology Co., LTD, China) in 0.5 mL of PBS plus 10 μg cholera toxin (CT, List Biological Laboratories, Campbell, CA, USA) as an adjuvant from week 1 through week 5 and oral 100 mg whey containing 10 μg CT from week 6 through week 7. Mice were then sensitized via intraperitoneal injection (i.p.) with 500 μg whey in 0.2 mL of PBS containing 2 mg Alum (ThermoFisher, USA) from week 8 through week 9. Naive mice received PBS alone. Sham mice received CT and alum alone. A schematic for the sensitization and challenge timeline was depicted in Figure 1. …

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

… For macrophage ablation, CD11b-DTR mice received a tail vein injection of 5µg/kg diphtheria toxin (DT, cat. #150, List Bio, Campbell, CA) …

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

Reagents

… Shiga toxin 2 from Escherichia coli was obtained from List Labs (Campbell, CA), …

• Product #162L – Shiga Toxin 2 from Escherichia coli

… Mice were injected intraperitoneally with 200 ng of pertussis toxin (List Biological laboratories, INC) on days 0 and 2 unless otherwise specified …

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 of experimental autoimmune myositis

… BALB/c mice were each immunized intracutaneously with 50-100 μl of an emulsion into three to four locations (a total of 200 μl) on the back on days 0, 7, and 14. One hour after the first immunization, pertussis toxin (500 ng in 100 μl saline; List Biological Laboratories) was intraperitoneally injected into each animal. In the present study, all treated animals underwent successful EAM, defined by a significant increase in spleen weight.

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)

… For PTX experiments, cells were incubated with 10 µM PTX (List Labs) overnight (o/n) at 37°C. Light stimulation employed the Xenon white light flash lamp (1 mW nominal output power, 10 ms flash separation) of the microplate reader. …

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)

Experimental autoimmune encephalomyelitis (EAE) induction

Female C57BL/6 wildtype mice (Jackson Laboratories) or NG2creER:MAPTmGFP mice (8–12 weeks old) were injected subcutaneously with 200 μg MOG35-55 peptide (synthesized by Protein and Nucleic acid facility, Stanford University) emulsified in complete Freund’s adjuvant (CFA) (Thermo Fisher Scientific) containing 10 mg/ml of heat inactivated Mycobacterium tuberculosis H37RA (Sigma-Aldrich); injections were at one site into each hind flank. On the day of immunization and 48 h later, animals received intraperitoneal injections of 300 ng of pertussis toxin (List Biological Laboratories). …

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)

Cell isolation and propagation

The uterine horns were collected from one 8-week-old C57BL female mice (Wuhan University Center for Animal Experiment). The Animal Ethics Committee of Wuhan University Center for Animal Experiment approved this study. Cell isolation and culture procedures were carried out according to the previous studies with minor modifications [24, 25, 29, 30]. In brief, fresh murine uterus from one female mouse was dissected and the uterine horns were collected as illustrated in Fig. 1A. Uterine horns were minced into pieces and dispersed into single cells by digestion with collagenase (StemCell, Vancouver, BC, Canada) plus trypsin. The primary mouse endometrial epithelial cells were co-cultured with irradiated mouse fibroblast 3T3 cells (J2 strain) (YongTech, Shenzhen, China) in primary epithelial culture basic medium (PECBM). PECBM contains DMEM and nutrient F-12 Ham (3:1) (v/v) (Sigma-Aldrich), supplemented with 5% FBS (GIBCO), 2 nM triiodothyronine (Sigma), 0.5% insulin–transferrin–selenium reagent (Life Technologies), 5 μg/ml transferrin (Life Technologies), 10 ng/mL epidermal growth factor (Sigma), 0.4 μg/mL hydrocortisone (Sigma), 1 nM cholera toxin (List Biological Labs), 0.5 μg/mL amphotericin B (Fungizone; Bristol-Myers Squibb), 40 μg/mL gentamicin (Gentacin; Life Technologies), and 5 to 10 mol/L Y-27632 (Enzo Life Sciences). …

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