Bacterial Toxin Research Citations

TC DNA immunization by applying plasmid DNA onto a mouse skin area wherein the hair was depilated by warm-waxing:

… Two days after the plucking, mice were anesthetized again, and the plucked area was cleaned with 70% ethanol, hydrated for 20 min with warm water, and paper-dried. Plasmid DNA (pCMV- or pCI-neo-sOVA, 50 µg) admixed with 10 µg cholera toxin (CT) (List Biological Laboratories, Campbell, CA) was gently dripped onto the hydrated area using a pipette tip. …

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

Cell culture:

Human neonatal epidermal melanocytes (HEM1455) were obtained from Cell Applications (San Diego, CA). Melanocytes were grown in HAM’s F10 media supplemented with ITS premix (Becton Dickinson, Franklin Lakes, NJ), 12-O-tetradecanoylphorbol-13-acetate (Sigma-Aldrich, St Louis, MO), 3-isobutyl-1-methylxanthine (Sigma-Aldrich), cholera toxin (List Biological Laboratories, Campbell, CA), …

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

Axon tracing:

Retrograde neuronal tracing was performed via injection of cholera toxin Beta-subunit (CTB) following injury. Four days prior to euthanizing, the mice were re-anesthetized, with a constant delivery of isoflurane, and the right sciatic nerve was exposed. Two microliters of a 1.5% solution of CTB (List Biological Laboratories) was injected into the nerve using a Hamilton syringe. The solution was slowly injected over a 1 min period and the syringe was withdrawn over an additional 2 min to prevent reflux. The muscle and skin were then sutured. Spinal cords were isolated, and cryosectioned at 18 m thickness. Alternating sections were stained against CTB via goat anti-CTB genotoxiod (List Biologicals) and detected with a fluorescently conjugated secondary. The ImageJ software (NIH) was used to measure neurite length. Neurite length was measured and plotted as distance from injury epicenter (0 m).

Immunohistochemistry:

Terminally anesthetized mice were perfused trans-cardially with PBS, followed by 4% paraformaldehyde in PBS (pH 7.4). Spinal cords were isolated, post fixed for 1 h and cryoprotected in 30% sucrose overnight. 18 m frozen sections were prepared, blocked for 1 h with 5% serum and then incubated with primary antibodies (rabbit anti-IBA-1 1:500, Wako; rabbit anti-GFAP 1:500, Dako; mouse anti-APC (CC1), 1:100, Millipore; rabbit anti-NG2, 1:500 a generous gift from Dr. Joel Levine; goat anti-CTB, 1:5000, List Biologicals; mouse anti-CD3, Dako) overnight at 4 C. …

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)

Materials:

VAMPtide substrates (Products #540, #541 and #542), BoNT/B light chain (LC ) (Product #620A), and BoNT/B holotoxin (Prod #136A) are products of List Biological Laboratories, Inc.

• Product #540 – VAMPtide Peptide Substrate (o-Abz/Dnp) Substrate for C. botulinum Type B Neurotoxin
• Product #541 – VAMPtide Peptide Substrate (FITC/DABCYL) Substrate for C. botulinum Type B Neurotoxin
• Product #542 – VAMPtide (Pya/Nop) Peptide Substrate for Botulinum Neurotoxin Type B
• Product #620A – Botulinum Neurotoxin Type B Light Chain, Recombinant
• Product #136A – Botulinum Neurotoxin Type B, Nicked, from Clostridium botulinum

Methods – The above materials were used in the following (refer to poster for details):

• Fluorimetric assays
• Sensitivity and LOD

Reagents:

Lipopolysaccharide (LPS) (Cat #: 421), … were purchased from List Biological Laboratories, Inc. (Campbell, CA), …

Results – Production of ROS is increased in cells cultured in a serum-poor medium and leads to enhanced propensity of cells to be activated by SFAs:

In our previous studies demonstrating that SFAs activate TLR2 and TLR4, cells were cultured in the serum-poor (0.25% FBS) medium. Thus, we examined how the responsiveness of cells to C12:0 or TLR agonists is affected by different serum concentrations in the culture media. The responsiveness of RAW 264.7 cells to C12:0, C16:0-BSA, or TLR agonists (LPS, a TLR4 agonist; Pam3CSK4, a TLR2 agonist) was greatly potentiated when the cells were cultured in the serum-poor (0.25%) medium compared with the cells cultured in 10% FBS ( ) …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

Adoptive transfer myocarditis:

<... Each mouse additionally received pertussis toxin (PT, 100 ng; List Biologicals, Campbell, California) on days 0 and 2 posttransfer as described previously [19,27,28]. The animals were euthanized on day 21 posttransfer, and hearts and pancreases were collected for histological evaluation of inflammatory changes. …

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)

Animals and induction of EAE:

… Rag^/ mice were sublethally irradiated and reconstituted with Bat3^+/ or Bat3^/ fetal liver hematopoietic cells as described⁴². EAE was induced by subcutaneous injection of mice with 100 µl of an emulsion containing CFA, 250 µg Mycobacterium tuberculosis extract H37-Ra (Difco), and either 100 µg MOG₃₅₅₅ (MEVGWYRSPFSRVVHLYRNGK) or 100 µg PLP₁₃₉₁₅₁ peptide (HSLGKWLGHPDKF; QCB BioSource International). Mice received 200 ng pertussis toxin (List Biological Laboratories) intraperitoneally on days 0 and 2. For cell transfer experiments, 2D2 Th1 were retrovirally transduced and sorted on GFP⁺ 5 days after initial stimulation, 210⁶ GFP⁺ cells were transferred i.v. to each Rag^/ recipient. All animals received 200 ng of pertussis toxin (List Biological Laboratories) i.p. at d0 and d2. Animals were monitored daily for development of EAE according to the following criteria: …

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)

CTB retrograde tracing:

Four weeks after SCI, animals were injected with 5 l of cholera toxin -subunit (CTB; 10 mg/ml; List Biological Laboratories, California). The rats were re-anesthetized by isoflurane inhalation (5% induction/1-2% maintenance) and an incision was made approximately over the left mid thigh. The proximal portion of the left sciatic nerve was exposed and injected with CTB utilizing a 10 µl Hamilton syringe.

Studies in unlesioned rats demonstrated that 4-7 days was sufficient for transport of CTB tracer through the dorsal columns to brainstem gracile nucleus termination …

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

Experimental animals and surgical procedures:

… After completion of the behavioral testing, the rats were reanesthetized with isoflurane, and the left median nerve was exposed and 5 µl of 5% cholera toxin subunit B (CTB, List Biological Laboratories, Campbell, CA) injected into the left median nerve. Seven days later tissue was collected for analysis. …

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

… Two monkeys received a single pressure-injection of 5 µl of unconjugated CTB (List Biological Laboratories, Campbell, CA) through a Hamilton syringe (5 mg/mL in 0.01 M Phosphate buffer, pH 7.5). …

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