Bacterial Toxin Research Citations

Experimental Protocols:

Mice were treated with an intraperitoneal injection of 1 mg/kg purified LPS from E. coli 0111:B4 (List Biologicals) 4 hr prior to intravital microscopy. The dose of LPS used in this study was sufficiently low that animals survived anesthesia and hepatic perfusion was not severely diminished. The purity of the commercial LPS was confirmed by inoculation into TLR4 knockout (TLR4^/) animals and observation of no measurable response in any of the parameters reported in this study (data not shown). For platelet depletion experiments, mice were injected i.p. with 50 l anti-mouse thrombocyte serum or 50 l control serum (adsorbed normal rabbit serum) 24 hr prior to LPS administration or E. coli infection. For neutrophil depletion experiments, mice were injected i.p. with 200 g anti-Gr-1 antibody or 200 g rat IgG control antibody 24 hr prior to LPS administration. Platelet and neutrophil depletion were confirmed with SD-IVM (see Figure 2). In some experiments, mice were treated with 2000 U DNase intravenously at various time points after infection with E. coli (noted in the figure legends). 

• Product #201 – LPS from Escherichia coli O111:B4

Mouse Treatments and Immunizations:

For immunization with OVA in complete Freunds adjuvant (OVA/CFA), OVA was emulsified in CFA at final concentration of 1 mg/ml OVA, and mice received hind footpad injection of 20 l of OVA/CFA as described (Webster et al., 2006).

Foxp3-DTR mice express simian diphtheria toxin (DT) receptor that allows for depletion of Foxp3⁺ regulatory T cells upon DT administration (Kim et al., 2007; Chinen et al., 2010). For regulatory T cell depletion, wild-type control or Foxp3-DTR mice were injected intraperitoneally with 1.5 g of DT … List Biological Laboratory, Campbell, CA, USA) on days 0, 1, 4, and 7 and examined on day 8. CD4⁺, CD25⁺, Foxp3⁺ regulatory T cells in lymph nodes were depleted by at least 12-fold by day 8.

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

… Frozen sections of the brain, the TG, and the eye muscles were processed for the immunocytochemical detection of the tracers cholera toxin subunit B and wheat germ agglutinin (goat anti-choleragenoid, 1:5000; List Biological Laboratories, Campbell, CA, USA; …

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

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