Bacterial Toxin Research Citations

Muscle afferent tracing:

Three days after PTx injury, animals were reanesthetized and muscle afferents of the deep radial nerve were labeled with 10 µl injections of a 1% solution of cholera toxin B (CTB) (List Biological Laboratories) into the extensor carpi radialis of both forelimbs. The solution was injected over 1 min, and the needle was withdrawn over an additional 1 min. The wound was closed by suturing the overlying muscle and skin. Preliminary data demonstrated that CTB transganglionically labeled both primary sensory afferents and retrogradely labeled motoneuron pools. Anterograde labeling is punctate. Whereas at high magnification we could see punctate ventral horn labeling in proximity to labeled motoneurons, we were not able to distinguish this labeling from other ventral horn label, including portions of motoneuron dendrites and labeled recurrent motoneuron axon collaterals. For this reason, we limited our quantitative CTB analysis to the dorsal horn and intermediate zone. To assay putative 1A boutons on motoneuron cell bodies and proximal dendrites, we examined VgluT1 presynaptic labeling. …

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

Mucosal Sensitization:

… Briefly, mice were administered 50 mg of OVA (Sigma-Aldrich, St Louis, MO) with 10 µg of CT (List Biologicals, Campbell, CA) in a final volume of 200 µl by using a mouse feeding needle 4 times a week for 4 weeks.

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

Injection of tracers:

For the immunohistochemistry experiments requiring retrograde tracing, six animals were anesthetized using 5% isoflurane with oxygen and placed in a stereotaxic apparatus (David Kopf Instruments, Tujunga, CA) and stabilized with non-perforating ear bars. … Two l of 1.0% solution of CTb (List, Campbell, CA) was slowly injected into the parabrachial nucleus over a period of 20 minutes, followed by a waiting period of 10 minute before the micropipette was retracted from its position to minimize leakage of the tracer. CTb was injected seven days prior to sacrificing the animals.

Confocal microscopy:

… The sections were incubated in 3 different cocktails: #1) rabbit anti-CGRP at a 1:200 dilution (Sigma, St Louis, MO) and lectin IB4 conjugated to AlexaFluor 568 at a 1:200 dilution (Molecular Probes); #2) rabbit anti-CGRP and guinea pig anti-P2X3 at a 1:25,000 dilution (Neuromics, Edina, MN); #3) goat anti-CTb at a 1:5000 dilution (List Biological), …

Author did not specify which CTB was utilized.  List Labs provides Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae)  and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt).

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

EAE induction and clinical scoring:

Age- and sex-matched mice were immunized subcutaneously with 100 µg MOG35-55 (Genway, China) in IFA (incomplete Freund’s adjuvant) supplemented with 500 µg Mycobacterium tuberculosis on day 1. On days 1 and 3, mice were injected with 300ng 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)

… Mice additionally received an intraperitoneal injection of 500 ng pertussis toxin (List Biological Laboratories, Campbell, CA) dissolved in 0.2 mL PBS on days 0 and 2 of the study. …

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)

 Table 1. List of immunological reagents, source and concentration

Injection of retrograde tracers:

Following recovery (710 days) from the NTS injection, animals were re-anaesthetised (as previously described) and either 1 l of 2% fluorogold (FG) or 2 l of 0.5% cholera toxin B subunit (CTB, Table 1) was pressure injected into the left side spinal cord intermediolateralis …

Biotin dextran aminecholera toxin B subunit for epifluorescence:

BDA sections were incubated in 10% NHS0.3% Triton-X-100 for 45 min, rinsed in Tris-buffered saline (TBS, 1  10 min) and then incubated in an appropriate streptavidin Alexa Fluor 594 or 488 for 2 h. The sections then had a further rinse in TBS followed by avidinbiotin blocking and incubation in goat anti CTB with streptavidin Alexa Fluor 594 or 488 … 

• Product #103 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

Transganglionic Labeling of Sciatic-Projecting Sensory Axons. Three days before sacrifice, sciatic nerves were injected bilaterally with a 1% (wt/vol) solution of CTB (List Biological Laboratories). …

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

Induction and evaluation of EAE:

Female C57BL/6 mice (8-10 week old) were immunized subcutaneously in the dorsal flank with 150 µg of myelin oligodendrocyte glycoprotein peptides (MOG₃₅₅₅) emulsified in complete Freund’s adjuvant (CFA; Chondrex, Seattle, WA, USA) on days 0 and 7. The mice were injected intraperitoneally (i.p.) with pertussis toxin (List Biological Laboratories, Campbell, CA, USA) at a dose of 500 ng per mouse on days 0 and 2, and at a dose of 200 ng per mouse on day 8. …

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)

Methods:

Recombinant GST-SV2c (List Prod #690) and recombinant GST Synaptobrevin-2 (List Prod #510A), a control protein, were attached to glutathione coated magnetic beads. Each set of coated beads was exposed to varying amounts of BoNT/A in buffer, washed, and incubated with SNAPtide (List Prod #520), a quenched fluorogenic (FRET) substrate. Detection of cleaved peptide was monitored using reverse phase HPLC with fluorescence detection.  

• Product #690B – Receptor for Clostridium botulinum Type A Neurotoxin, GST-SV2c luminal domain
• Product #510A – Recombinant GST Synaptobrevin-2 (PRODUCT DISCONTINUED)
• Product #520 –  SNAPtide Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin

TNA:

The titer of toxin-neutralizing antibody in serum was determined using modifications to the methods of Pitt et al [15] and Little et al [4] by testing the ability of the serum to inhibit the cytotoxicity of the combination of rPA with LF (List Biological Laboratories Inc., Campbell, CA, USA). The assay was carried by exposing J774A.1 murine macrophages (ATCC TIB-67) to PA and LF in the presence or absence of serum. In brief, J774A.1 cells were plated out on 96 well plates (SPL CO. Ltd, Pyeongtaek-si, Korea) and seeded with 10⁶ cells per well in 150 l volumes 2 hours before testing. …

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