Bacterial Toxin Research Citations

Single molecule fluorescent in situ hybridization (RNAscope) and immunohistology

… Immunohistology was performed as described with minor modifications.89 In short, sections were thawed, briefly washed (PBS with 0.2% Triton X-100), and blocked (PBS with 0.2% Triton X-100 and 5% normal horse serum) for 1 hour at room temperature. The primary antibody was diluted in blocking solution and incubated for 1-2 days at room temperature. Sections were washed in PBS before being incubated with the secondary antibody diluted in blocking solution for 1 hour at room temperature. Sections were again washed in PBS and mounted with Immu-Mount (ThermoFisher). The following primary antibodies were used: goat anti-Phox2b (R&D Systems, AF4940, 1:200), rabbit anti-RFP (Rockland, 600-401-379-RTU, 1:500), chicken anti-GFP (Aves Labs, GFP-1020, 1:500), rat anti-GFP (Nacalai Tesque, GF090R, 1:1000), goat anti-CD117 (R&D Systems, AF1356, 1:400), rabbit antiTubulin b-3 (BioLegend, Poly18020, 1:1000), goat anti-CTb (List Labs, 703, 1:2000), …

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

… Control and immunised mice received 200 ng pertussis toxin (List Biological, California, US) by intraperitoneal injection on the immunisation day and 2 days post …

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)

Induction of EAE

12-week-old male hu-TIGIT KI mice were subcutaneously (s.c.) injected with 200 µg of MOG35–55 peptide (Synpeptide, Shanghai, China) and 400 µg of Mycobacterium tuberculosis in incomplete Freund’s adjuvant (Difco Laboratories, NJ, USA). Pertussis toxin (400 ng per mouse; List Biologicals, CA, USA) was intraperitoneally injected the same day and on day 2 post immunization. We also intraperitoneally administered the anti-hu-TIGIT agonistic mAb (100 μg per mouse in 100 μl PBS) or a mouse IgG1κ isotype control (clone MG1-45, BioLegend, 100 μg per mouse, in 100 μl PBS) to each mouse on days 0, 2, 4, 10, and 17. …

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)

… Diphtheria toxin (DT) List Biological Laboratories Cat#150 …

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

Immunohistochemistry:

… Some of the brains (n=3) from WT mice were injected with CTb and were immunostained using Goat anti CTb (1:30K, Cat# 703, RRID: AB_10013220, List Biological Laboratories Inc., CA). Sections for double staining for GFP, mcherry, CTb or FoxP2 were incubated in fluorescent-labeled secondary antibodies (Alexa- 488 at 1:200 or Alexa- Cy3 at 1:200; …

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

2.2. Wt-RABV injections in the inferior live of the rat (n = 10)

The wt-RABV stock was kept at −80◦C until use. The injectate consisted of a mixture of 4 parts wt-RABV and 1 part cholera
toxin β-subunit [CTb, low salt; List Biological Laboratories, 1% w/v in 0.2 M phosphate buffer (PB) at pH 7.4]. CTb was added to accurately evaluate the injection site and to be able to discriminate between non-transneuronal (CTb) and non-transneuronal and transneuronal infection by the wt-RABV (Ruigrok et al., 2008; Suzuki et al., 2012). Injection of approximately 150 nl was made with a 1 µl Hamilton needle attached to a microsyringe pump (Ruigrok et al., 2016). The IO was reached at a −45◦ angle with the vertical axis, with the needle entering at 0.6–0.8 mm lateral to obex at a depth of 2.7–2.8 mm. After injection the neck muscles were sutured in layers, the skin was clamped and the animal was allowed to recover. …

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

… Prediluted serum samples (1:50) were diluted by 2-fold in another 96-well plate and incubated with a constant amount of anthrax lethal toxin (100 ng/mL PA and 80 ng/mL LF; List Biological Laboratories, Campbell, CA, USA) for 30 min. The generated toxin-antiserum was then applied to J774A.1 cells and incubated for 4 h. …

• 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:

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

Stereotaxic surgery.

… For retrograde tracing of TRN-projecting and PVT-projecting PL cells CTB-488 and CTB-555 (1.0% in PBS, Thermo Fisher Scientific) were injected into the TRN (0.5 µL) and PVT (1.0 µL), respectively, and allowed 4 d for retrograde transport. For retrograde tracing of PVT-projecting TRN cells unconjugated CTB (List Labs Product No. 104) was injected into the PVT (1.0 µL). For retrograde labeling of PVT-projecting TRN cells for RNAscope, retrobeads (LumaFluor, Inc.) were injected into the PVT and allowed 7 d for retrograde transport. …

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

… immunohistochemistry. Briefly, sections … were processed and incubated overnight in a goat anti-CTb antibody (List Biological Lab, #7032A6) at 1:10,000, followed by rinses in PBS and incubation …

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

Induction of EAU

To effectively induce EAU production, we injected mice subcutaneously with a 1:1 mixture of 200 µg IRBP1−20 (GL Biochem, Shanghai, P. R. China) and complete Freund’s adjuvant (Difco, Detroit, MI, USA) including 2.5 mg/mL of Mycobacterium tuberculosis strain H37Ra (BD Difco, San Jose, CA, USA). After immunization, 0.25 μg of PTX (List Biological Laboratories, Campbell, California, USA) was injected intraperitoneally 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)