Bacterial Toxin Research Citations

Experiment 3: Retrograde Tracing from the SCN to the Forebrain During WC. – Cholera toxin administration:

The hamsters were positioned in a stereotaxic frame with the incisor bar set to 2.0 mm below the interaural level. The scalp was incised and a burr hole drilled at 0.5 mm anterior, 1.25 mm lateral from bregma. A glass electrode was back filled with the subunit of cholera toxin (CT; 1% in distilled H₂O; List Biological Laboratories). A wire composed of 10% (wt/wt) iridium and platinum was placed in the CT and attached to the lead, …Once lowered, the CT iontophoresis took place using a positive current of 1.5 A in alternating 5 s on/5 s off pulses for 10-12 min. Following the iontophoresis, …

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

EAE:

… In brief, B6 mice were s .c. immunized with 100 µg of MOG 35 55 peptide in complete Freund’s adjuvant containing 0.4 mg Mycobacterium tuberculosis H37RA (Difco, Lawrence, KS). Two hundred nanograms of pertussis toxin (List Biological Laboratories, Campbell, CA) was adminis tered i.v. on day 0 and day 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)

Sensitization and challenge:

Mice were gavaged two times at a weekly interval with 2 mg of food proteins or 20 mg WPI in PBS with 10 µg cholera toxin (CT; List Biological Laboratories, Inc., Campbell, CA). Control animals received PBS and CT only. …

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

Materials:

… PA from B. anthracis in its holo form (PA83) and cleaved form (PA63) where purchased from List Biological Laboratories, …

In vitro selection scheme:

The AEGIS-LIVE experiment was performed on a synthetic library of GACTZP oligonucleotides containing 25 randomized positions flanked by two primer binding sites (59 nt in length, 5-AGAGAGCGTCGTGTGGA-N25-TGAGGAGGTGCGCAAGT-3).

PA was presented immobilized on magnetic beads…

Filter binding assays:

5-³²P-labeled aptamer (1 l each) was denatured/renatured in 1 PBS (30 l), and incubated (45 min, RT) with increasing concentrations of PA63 or PA83, from 0 to 12 M. To capture the PA and PA-aptamer complex, aliquots (5 l) from each binding reaction were spotted on a nitrocellulose membrane on top of a charged nylon membrane (Whatman) in a dot-blot minifold vacuum device (Millipore) (27). Membranes were washed …

Determination of the affinity constant with bead-based assay:

After being coupled to PA63 (1.6 nmoles), the beads (200 l, as during selection) were divided into aliquots (20 l each) and each fraction was incubated with increasing concentrations of …

Enzymatic footprinting:

For DNase I digestion of PA1 and PA1T4 in the presence of PA63 or PA83, the aptamer was denatured (85C, 5 min) followed by slow (0.1/s) cooling to 24C to allow refolding. 5³²P-PA1 with 05 M PA63 or PA83 was incubated with tumbling for 45 min, and then treated with DNase I nuclease (15 min at RT). The digested products were resolved on 18% denaturing PAGE. …

Electrophysiology assays:

LF N-terminal domain (LF_N) and PA were expressed and purified as described (21). To form heptameric PA oligomers, PA was nicked with trypsin (1:1000 w/w) at room temperature in buffer (20 mM TrisCl, pH 8) …

• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis
• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

… For alloreactive T cell-proliferation studies, the plates were incubated at 37C in a 5% CO 2 humidified incubator for 5 d. For microbial Ag-specific T cell-proliferation studies, the plates were incubated for 7 d. TT from Clostridium tetani (List Biological Laboratories), …

• Product #190 – Tetanus Toxin from Clostridium tetani

EAE Induction:

Female SJL mice (612 wk old) were obtained from Jackson Laboratories. EAE was induced by s.c. injecting 200 µg proteolipid protein (PLP) peptide …Two hundred nanograms of pertussis toxin (PTx) (List Biological Laboratories) dissolved in 200 µL PBS was administered 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)

Reagents:

PA₆₃ was purchased from List Biological Laboratories, Inc., (Campbell, CA, USA). …

Channel Reconstitution into Planar Lipid Bilayers:

To form solvent-free planar lipid bilayers with the lipid monolayer opposition technique [81], we used …Single channels were formed by adding 0.5 to 1 L of 20 gmL¹ solution of PA₆₃ to the 1.5 mL aqueous phase in the cis-half of the bilayer chamber. Under this protocol, PA₆₃ channel insertions were always directional, as judged by channel conductance asymmetry in the applied transmembrane voltage. …

• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Table 3.2:

Primary antibodies for in vivo growth in SC sections

Primary antibody Made in  Producer Dilution used

Anti CTB Gt List Biological 1:80000

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

Reagents:

LPS was obtained from List Biological Laboratories (San Jose, CA, USA). …

Author did not specify which List Labs LPS product was utilized in their research.  List Labs provides the following LPS products:  https://listlabs.com/product-information/lipopolysaccharides/

Materials:

…while ETA, DT, and CT were purchased from List Biologicals (Campbell, CA). …

CT toxicity assay:

CHO-K1 cells (ATCC #CCL-61) were co-incubated with a combination of CT and grape compound for 18 h before cAMP levels were quantified as previously described…

Vero-d2EGFP toxicity assay:

Ricin, ETA, DT, and ST1/ST2 intoxication assays were peformed with Vero-d2EGFP cells as previously described…

Author did not specify which Cholera toxin was utilized.  List Labs provides Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae and Product #101 – Cholera Toxin from Vibrio cholerae.

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #160 – Exotoxin A from Pseudomonas aeruginosa