Bacterial Toxin Research Citations

Immunofluorescence labeling of myelinated afferent fiber projections to the dorsal horn:

To detect the effect of -FNA on RTX-caused myelinated primary afferent nerve sprouting into the spinal dorsal horn, cholera toxin B-subunit (CTB), a transganglionic tracer, was injected into left sciatic nerve of VEH-, RTX-, EA + NS- and EA + -FNA-treated rats as described.4 The results were analyzed using immunofluorescence labeling.

In brief, rats were deeply anesthetized with sodium pentobarbital (40 mg/kg, i.p.) and transcardially perfused 4 days after CTB injection. The L4-L6 spinal cord was post fixed in 4% paraformaldehyde for 68 hr and dehydrated in 30% sucrose dissolved in 0.1 M phosphate buffer (PBS) at 4 C for 48 hr. The tissues of spinal cord were sectioned with a cryostat. The thickness of sections was 20 m. The slides were incubated in blocking buffer (0.2% tween-20 and 5% donkey serum in 0.01M PBS, pH 7.4) at 37 C for 1 hr, then incubated overnight on a shaker at 4 C with primary antibodies diluted in 0.01M PBS including 0.3% TritonX-100, 5% donkey serum, and incubated with a secondary antibody at 37 C for 1 hr. The primary antibodies used were goat anti-CTB (1:250; List Biological Laboratories, Campbell, CA). For secondary antibodies, we used donkey anti-goat IgG conjugated with Dylight 594 (1:500; Jackson ImmunoResearch). We used 6 rats in each group and randomly selected five or six sections from the spinal cord of each rat. All imaging analysis was performed on an Olympus BX51 fluorescence microscope (Olympus, Tokyo, Japan) and using Image J software (Bethesda, MD).

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

… tracer Cholera toxin B fragment (CTB low salt, end concentration 0,03%) (List Biological Labs, Campbell, CA) was injected in a single dose (2 l delivered over 30 minutes) into was visualized using as primary antibody goat anti-choleragenoid (List Biological Labs …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

EAE development and anti-IL-17 antibodies treatment:

… EAE was induced in 9-week-old female C57BL/6 mice. Mice were immunized subcutaneously with an emulsion containing myelin oligodendrocyte glycoprotein(MOG) 3555 peptide (synthesized in-house) and Complete Freund’s Adjuvant (CFA). Immunization was performed under isoflurane anesthesia. All animals received pertussis toxin (List Biological Laboratories, Campbell, CA) in PBS intraperitoneally (i.p.) on day 0 and day 2 after MOG immunization. …

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 also received 400 ng of pertussis toxin (LIST Biological Laboratories) intraperitoneally both immediately after immunization and 48 h later. Control mice were injected with everything except the MOG peptide …

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)

Recombinant antigens and coupling to microsphere beads:

… Tetanus toxoid (Massachusetts Biological Laboratories, Boston, MA), diphtheria toxoid from Corynebacterium diphtheriae (List Biological Laboratories, Campbell, CA), and recombinant measles nucleoprotein (MV-N, Meridian Life Sciences, Memphis, TN) [30] were purchased from commercial sources. Tetanus toxoid was coupled to SeroMap beads as previously described [31]. Diphtheria toxoid was coupled in buffer containing 50 mM 2-(N-morpholinoethanesulfonic acid (MES) at pH 5.0 with 0.85% NaCl at a concentration of 60 g of protein per 1.25 10⁷ beads in 1 ml final volume. …

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

… Diphtheria Toxin (unnicked) List Biological Laboratories Cat # 150 …

• Product #150 – Diphtheria Toxin, Unnicked, fromCorynebacterium diphtheriae

LPS:

Purified LPS from Salmonella Typhimurium (catalog no. 225) was obtained from List Biological Laboratories, Inc. (Campbell, CA) …

• Product #225 – LPS from Salmonella typhimurium

… Pertussis toxin was purchased from List Biological Laboratories (Campbell, 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)

… Cholera toxin was from List Biological Laboratories (cat. no. 100B). …

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

Lactate dehydrogenase (LDH) assay:

Macrophages were pretreated for 30min with glycine or one of the other small-molecule inhibitors at the indicated dose. All chemicals were purchased from Sigma-Aldrich, with the exception of propofol (EMD Millipore). Late-log cultures of Salmonella typhimurium SL1344 grown in L-broth containing 0.3M NaCl were used to infect macrophages (MOI 10:1) for 90min, unless otherwise indicated. Cells were treated with anthrax lethal toxin comprised of 1g/ml protective antigen and 1g/ml lethal factor (List Biological) for 2h¹⁴. Cells were treated with the detergent-based lysis buffer included in the Cytotox 96 Kit (Promega) for 30min. Release of cytoplasmic LDH was determined from triplicate samples using the Cytotox 96 Kit and calculated as 100 (experimental LDHspontaneous LDH)/(maximum LDHspontaneous LDH).

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

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Product #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence