Bacterial Toxin Research Citations

Materials:

Hinute-AM was kindly provided by Fuji Oil Co., Ltd. We used smooth-type LPS from E. coli O55:B5 (List Biological Laboratories, CA, USA) as …

NOTE:  At the time this paper was written, Product #203 (5mg – LPS from Escherichia coli O55:B5) may have been utilized.

Product #203 (5 mg – LPS from Escherichia coli O55:B5) is no longer sold.

Product #203A (2.5 mg – LPS from Escherichia coli O55:B5) is available for purchase.

• Product #203A – LPS from Escherichia coli O55:B5

… Additionally, the mice receive a peritoneal injection of 200 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA) dissolved in PBS …

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)

Protease assay:

Proteolytic activity was determined, as described earlier [22Kaman WE, Hulst AG, van Alphen PT, et al. Peptide-based fluorescence resonance energy transfer protease substrates for the detection and diagnosis of Bacillusspecies. Anal Chem. 2011;83:25112517.[Crossref], [PubMed], [Web of Science ], [Google Scholar]], by incubating 16 M FRET-peptide with varying concentrations of PA₈₃ or LF (both List Laboratories) in HEPES buffer (20 mM HEPES, 0.05% Tween-20, pH 8.2) using blackwell clear bottom 96-well plates (Corning). …

Cleavage characteristics of PA₈₃

20 g/mL PA₈₃ was incubated with 16 M FRET-peptide PEK-054 (FITC-NleKKKKVLPIQLNAATDK-KDbc) [20Cummings RT, Salowe SP, Cunningham BR, et al. A peptide-based fluorescence resonance energy transfer assay for Bacillus anthracis lethal factor protease. …

LC MS/MS:

Prior to LC MS/MS analysis 10 g PA₈₃ was digested with 0.5 g trypsin (Difco, BD Biosciences) for 3 hr at 37C. The trypsinized sample was analyzed by MS/MS using the data dependent LC-auto MS/MS mode. …

SDS-PAGE:

PA₈₃ protein (2 g) was mixed 1:1 with 2x Laemmli sample buffer (Biorad), incubated for 5 min at 95C and loaded onto a 10% SDS-PAGE gel. After electrophoresis for 1 hr at 22 mA (max. 160 V) the gel was stained overnight with PageBlue staining solution (Fisher Scientific) and washed two times to visualize bands.

Specificity testing PA₈₃

20 g/mL PA₈₃ was incubated with 15 g/mL B. anthracis protective antigen antibody-[HRP] (-PA, Novusbio, BAP0102), 15 g/mL human IgG antibody-[HRP] (-human IgG, Sigma Aldrich, A0170) or PBS (solvent of the used antibodies) in HEPES buffer for 5 hr at room temperature. Next, 1 L PEK-054 (800 M) was added to the reactions and increase in fluorescence was measured at 37C for 60 min as described above.

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

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

… Bordetella pertussis toxin is given List biological laboratories Inc (Campell, 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)

… 200 ng of pertussis toxin (PTX) (List Biological Laboratories) was injected intraperitoneally on the same day as MOG immunization and again two days later. …

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)

… Injection of retrograde tracer into preBotzinger complex In three animals, preBotC injections of the retrograde tracer, Cholera Toxin b (CTb, 1% in deionized water; List Biological, Campbell, CA) were unilaterally performed. …

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

Peptide:

Tetanus Toxoid (Lot #19119A1) List Biological Laboratories, Campell, USA

ELISpot Assay:

… As negative controls served four cell suspension containing
Holes without further additions and two holes each, in which DMSO in the same
Dilution as the dissolved in DMSO peptides were added. As positive
Control antigens were tetanus toxoid at a concentration of 0.3 g / ml and
Phytohemagglutinin (PHA) at the concentration of 3 g / ml. …

• Product #191 – Tetanus Toxoid from Clostridium tetani

For retrograde tracing, 1 µl of cholera toxin B subunit (CTB) solution (10% dilution with ddH2O; List Biological Laboratories), …

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

INDUCTION OF EPILEPSY: TETANUS NEUROTOXIN INJECTION:

… Each animal was anaesthetised and placed into a stereotaxic frame. 2.5 ng TeNT (List Biological Laboratories Inc., United States) in 1 L of 2% bovine serum albumin in phosphate buffered saline was injected through the guide cannula, at 200 nL min-1 using a Hamilton Syringe (25G, 7001 KH, Hamilton Company, United States) and a microinjection pump (WZ-74905-02, Cole-Parmer, United Kingdom).

• Product #190 – Tetanus Toxin from Clostridium tetani

… Goat anti-CTb, List Biological Labs, Cat#703. …

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