Bacterial Toxin Research Citations

… Diphtheria toxin List Biological Laboratories, Inc Un-nicked, Lot #: 15043A1 …

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

… Nicked BoNT/E3 was obtained from List Biological Laboratories (Campbell, CA, USA). …

Sandwich Assay Procedure:

When performing the sandwich assay, for every mAb, the unlabeled mAb was used as the capture antibody and the biotinylated form was used as the detector antibody. Each mAb was tested in combination with all of the mAbs to identify pairs that can detect the BoNT/E3 toxin in the solution phase. Black plates were coated with 100 L of mAb at 2 g/mL in 0.05 M sodium carbonate buffer, pH 9.6, incubated overnight at 4 C, and then blocked with 3% NFDM-TBST. The blocking solution was removed and BoNT/E3 toxin in 3% NFDM-TBST was added at an initial concentration of 500 ng/mL and serially diluted two-fold, including wells with no toxin. Next, biotinylated-mAb was added in triplicate at 1 g/mL, 100 µL/well in 3% NFDM-TBST. Plates were incubated for 1 h at 37°C, washed 6 with TBST, and visualized using SuperSignal West Dura Extended Duration Substrate as described above.

When the most sensitive combination of capture mAb and detector mAb-biotin were selected, the final sandwich assays were performed in triplicate using BoNT/E3 holotoxin at a starting concentration of 1000 ng/mL, serially diluted three-fold, including wells with no toxin, and visualized using SuperSignal Femto Max Sensitivity substrate. Limit of detection cut-off values were determined using three times the standard deviation of the wells with no toxin.

EAE induction and clinical scoring:

… Pertussis toxin (500ng/mouse, List Biological Laboratories, Inc., Campbell, CA) was injected intraperitoneally (i.p.) on days 0 and 2. Animals were monitored daily for EAE signs as described⁵⁶.

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)

… Goat anti-CTb (1:5000) List Biological Laboratories Inc., CA, USA. Cat#703; …

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

C. difficile cytotoxin assay:

Cytotoxicity assay was performed as previously described with the following modifications 189. Briefly, ATCC CCL-81 Vero cells were grown to confluence in Dulbecco modified Eagle medium (Gibco 11965) supplemented with 10% fetal bovine serum (16140) and 1% Penicillin streptomycin (Gibco 15070). Cells were plated to a density of 105 cells/well. Mouse cecal content was diluted 1:10 in sterile PBS, passed 46 through a 0.22m filter, and serially diluted to 10-6 . Filtered samples were tested in duplicate with a corresponding control which both antitoxin (Techlab T5000) and sample were added. A positive control of C. difficile TcdA (List Biologicals 152C) was used. Samples were incubated overnight at 37C and the cytotoxic titer was determined as the reciprocal of the highest dilution that produced 80% cell rounding.

• Product #152C – Toxin A from Clostridium difficile

Cell Culture:

… cholera toxin (100 ng/ml; List Biological Laboratories …

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

Test substances:

… Clostridium difficile Toxin A (ToxA) was purchased from List Laboratories, Inc. (Campbell, CA). …

Evaluating endpoint intestinal monolayer barrier integrity and IEC viability:

… Cytotoxic responses were also examined for BSA, ToxA, and WGA along with positive and negative controls. As expected, TX-100 caused complete loss of T84 monolayer viability even upon a single exposure. EDTA and BSA did not elicit any cytotoxicity, nor did a single exposure to WGA or ToxA (Fig. 5H). Repeated exposures to WGA caused a significant degree of loss in MTT conversion indicating loss of cell viability. Repeated exposures to ToxA caused complete loss of T84 monolayer viability, a strikingly different response than observed with a single exposure to ToxA, which elicited no change in T84 monolayer viability (Fig. 5H). …

• Product #152C – Toxin A from Clostridium difficile

… Along with immunization and two days post immunization, 100 ng pertussis toxin (List-Biological-Laboratories) in Dulbecco’s phosphate buffered saline (DPBS; Sigma Aldrich) was administered by intraperitoneal injection …

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)

 EAM Induction

To induce EAM, we injected mice with 125 g myosin heavy chain peptide MyHC_614-629 (Ac-SLKLMATLFSTYASAD; Genscript) (Pummerer et al., 1996) emulsified in CFA (Sigma-Aldrich) supplemented with 5mg/mL heat-killed Mycobacterium tuberculosis strain H37Ra (Difco) on days 0 and 7. On the first day of immunization, mice also received a dose of 500 ng pertussis toxins intraperitoneally (List Biological Laboratories) (Cihkov et al., 2004).

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)

… Cholera toxin (Lot#10067A1) was obtained from List Biological Laboratories, Inc. (Campbell, USA. …

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