Bacterial Toxin Research Citations

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

… biological sample (S. aureus) Staph enterotoxin B List Biological Laboratories #122 …

Mixed lymphocyte cultures:

Mouse splenic CD4⁺ T cells were purified with a Stem Cell Technologies negative selection CD4⁺ T cell isolation kit and co-cultured with bone marrow-derived dendritic cells (cultured with GM-CSF for 7 days) at 1:1 ratio with or without Staphylococcal entertoxin B (SEB, 0.05 g/ml, List Biological Laboratories,>95% pure, 14 EU/mg) and in the presence of either IL-21R-Fc blocking agent or control Fc (20 g/ml) for 24 hr; RNA was then isolated. For intracellular staining of IL-21, human CD4⁺ T cells were purified from peripheral blood (Dynabeads Untouched negative selection CD4⁺ T cell kit, Invitrogen) and co-cultured at a 5:1 ratio with monocyte-derived DCs that had been cultured for 6 days in GM-CSF (800 U/ml) plus IL-4 (400 U/ml). At 72 hr, cultures were treated with Golgi Plug for 4 hr and stained 

… Fourteen days post sciatic nerve crush sensory axons that had re-innervated the muscle were traced by injecting 5 µl of 1% CTB (List Biological Laboratories) diluted in saline into the tibialis anterior and medial gastrocnemius muscles using a 10 µl Hamilton syringe and Hamilton needle (NDL small RN ga34/15mm/pst45o ) (Hamilton). …

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

Engineering cell sheets with oral mucosal epithelium cells on transwell:

The isolated epithelial cells were seeded on sixwell plate Transwell (Corning, Tewksbury, MA), at 65,000 cells/cm² in coculture with mitomycin C (MMC)treated NIH/3 T3 feeder cells (Hayashida et al., 2005). OMECs were seeded in the Transwell (Corning, Tewsbury, MA) and were cultured with MMCtreated NIH/3 T3 feeder cells in the following culture media: The culture medium was a mixture of Dulbecco’s modified Eagle’s medium (Sigma, St Louis, MO) and Ham’s F12 (Sigma, St Louis, MO) at a ratio of 3:1 supplemented with 10% fetal bovine serum (ThermoFisher Scientific, Chino, CA), 0.4 g/ml hydrocortisone (Sigma, St Louis, MO), 2 nmol/l triiodothyronine (Sigma, St Louis, MO), 1 nmol/l cholera toxin (List Biological Laboratories, Inc., Campbell, CA), …

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

Induction of EAE

EAE was induced by subcutaneous immunization with 50 µg of the myelin oligodendrocyte glycoprotein 3555 peptide (MOG₃₅₅₅; Peptide International, Louisville, KY, USA) emulsified in complete Freund’s adjuvant (Difco Laboratories, Detroit, MI, USA) supplemented with Mycobacterium tuberculosis extract H37Ra (Difco Laboratories) on Day 0. In addition, mice received 250 ng pertussis toxin (List Biological Laboratories, Campbell, CA, USA) intravenously on Day 0 and intraperitoneally on Day 2. Control mice were treated with complete Freund’s adjuvant and saline alone. EAE mice were divided into two groups based on the body weight, and treated vehicle or MR16-1.

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)