Bacterial Toxin Research Citations

smFISH and immunostaining

… For immunostaining, sections were permeabilized in PBS + 0.3% Triton X-100, blocked for 1 h in block buffer (PBS + 0.3% Triton + 10% normal donkey serum) and incubated with primary antibodies in block buffer at 4 °C overnight. Slides were washed three times, incubated in secondary antibodies in block buffer for 1 h at room temperature and washed three times, and a coverslip was applied with ProLong Gold Antifade Reagent. Primary antibodies included: chicken anti-GFP (Aves Labs, GFP-1010, 1:1,000), rabbit anti-c-Fos (Synaptic Systems, 226 003, 1:5,000), goat anti-ChAT (Millipore, AB144P, 1:100) and goat anti-CTB (List Labs, 703, 1:1,000). Species-specific donkey secondary antibodies conjugated to Alexa Fluor 488, 568 or 647 were obtained from Life Technologies or Jackson ImmunoResearch and used at a 1:500 dilution. …

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

… Animals were given two 400 ng intraperitoneal injections of B. pertussis toxin (Cat# 181; List Biological Laboratories) on days 0 and 2 …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

… 5 mg/mL heat-killed 493 Mycobacterium tuberculosis (Cat#231141, BD Bioscience), followed by a tail vein injection of 200 ng of pertussis toxin (Cat#179B, List Biological Laboratories) on day 0. Intraperitoneal injection of 200 ng of pertussis toxin was administered on day 2 …

• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

… DT, unnicked (List Biological Laboratories), was injected intraperitoneally at a dose of 20 ng per g of body weight in 1 × PBS for initial depletion and phenotyping. Where applicable, DT at a dose of 4 ng per g of body weight in 1 × PBS was then injected intraperitoneally every 3 d for continuous depletion. We used 1 × PBS alone as a vehicle control. …

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

Animal experiment

Animal experimental procedures were approved by the Animal Research Committee of Shandong University Affiliated Shandong Provincial Hospital. The C57BL/6J male mice weighing approximately 25 g were used in this study. … To induce EAE, MOG 35 to 55 peptide (Sigma-Aldrich) was mixed with CFA [incomplete free adjuvant (Sigma-Aldrich) and mycobacterium tuberculosis (BD Difco)], followed by subcutaneous injection of 100ul on each side of the lower back of 6-week-old male C57BL/6J mice. Pertussis toxin (List Laboratories) at a dose of 200 ng was injected intraperitoneally into the mice on days 0 and 2 after immunization. Mice were weighed daily and assessed for clinical symptoms. …

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)

Microglia depletion

For microglia depletion, 80 ng of diphtheria toxin (DT) (0.5 ng/μL, 150, List Labs, USA) was injected intraperitoneally on day -2 and 0 before SAH induction.

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

EAE induction

EAE was induced by immunization 8 to 10-week-old female C57BL/6J mice with 250 mg MOG33-55 peptide (Genemed Synthesis)
emulsified in complete Freund’s adjuvant (CFA) (BD Difco) per mouse, injected subcutaneously into each flank, followed by intraperitoneal injection of 150 ng of pertussis toxin (PT) (List Biological Laboratories, Inc.) per mouse on day 0. A second intraperitoneal injection of 300 ng of PT was administered on day 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)

… For EAE alone, mice were immunized with 100 µg of MOG35–55 (Genemed synthesis, Inc) in an emulsion containing complete Freund’s adjuvant (CFA, Difco), then injected peritoneally with 200 ng of Bordetella pertussis toxin (lot# 181236A1, List Biological Laboratories) on post-immunization day 0 and 2. For CPZ/EAE treatment, Abcd1y/− and wild-type mice were fed 0.2% CPZ diet for 2 weeks followed by MOG immunization18. We performed two separate rounds of CPZ alone and EAE alone treatments, and we also conducted CPZ/EAE combined treatment at three different time points. We used 2–3 mice per group in each round. …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)