Citations

… For optimum visualization of CTb, sections were incubated overnight at RT with a goat anti-CTb antibody (1:10,000 List Biological Laboratories, CA, USA) diluted in 0.1 M PBS containing 0.3 % Triton X-100 and 0.5 % BSA. …

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

Induction of EAE:

Induction of myelin oligodendrocyte glycoprotein (MOG) EAE was done as previously described.30,31 Shortly, 6- to 8-week-old female C57BL/6 mice were immunized with an emulsion containing 200 µg of MOG_3555…The inoculum (0.2 mL) was injected subcutaneously into right and left flanks. One hundred nanograms of pertussis toxin (List Biological Labs, Campbell, CA, USA) in 0.1 mL saline was also injected intraperitoneally on day 0 and 48 hours later.

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)

Analysis of TMV-PA Antigens for Reactivity against PA Specific Antibodies:

SDS-PAGE, ELISA and Western Blots – 
 
… For westerns, gels were transferred onto 0.2 um nitrocellulose
membranes (7 cm x 8.5 cm) in Tris-Glycine buffer (25 mM Tris, 192 mM Glycine, 20%
Methanol, pH 8.3) for 1 hour at 100V. Membranes were blocked with 2.5% non-fat milk
in TBS (20 mM Tris, 150 mM NaCl, pH 7.6) for 1 hour at room temperature. Primary
antibody (Goat anti-PA polyclonal, List Biologics Lot # 7712A2 …

… (c) Wash sections with 0.01 M PB three times for 5 min each time. (d) Bathe sections in primary anti-CTB antibody, for example, List (List Biological Lab) goat anti-CTB (1:5005,000) in 0.01 M PB containing 1 % BSA and 1 % TX-100 either overnight at 4 C or 35 h at RT. …

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

Induction of Active EAE in Mice by Immunization with MOG and Adjuvant:

EAE was induced in female wild-type C57BL/6 mice or MT and IL-10deficient mice on C57BL/6 background…All mice were given 400 ng of pertussis toxin (List Biological) intraperitoneally at 0- and 48-h postimmunization.

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:

68-week-old female C57BL/6 mice were immunized subcutaneously on day 0 with 75 µg/mouse of MOG3555 peptide emulsified in complete Freund’s adjuvant supplemented with 400 mg/ml Mycobacterium tuberculosis H37Ra (Difco). Each mouse was subsequently administered 250 ng pertussis toxin (List Biological Laboratories) intraperitoneal on days 0 and 2 after MOG₃₅₅₅ peptide 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)

Synthesis of cholera toxin-based vaccine

Cholera toxin (CT) was obtained from List Biologicals Laboratories (USA) …

Immunizations and injections of antibodies

C57BL/6 mice synchronized by Depoprovera® treatment were immunized under anesthesia by vaginal application of CTBOVA (4μg) + CT (1μg) in PBS at days 0, 14, and 21. NKp46+ ILC cells were depleted after intraperitoneal (i.p) injection of 200μL of anti-NK1.1 mAb (PK136 clone, 1/20 dilution of ascite fluid) at days -9 and 2 before each immunization. Such protocol insured at least a 98% depletion of NKp46+CD3- ILC cells in spleen, genital lymph nodes and vagina without affecting invariant natural killer T (iNKT) cells. NKG2D/NKG2D ligands blockade was performed by i.p injection of 500μg of mAb anti-NKG2D (CX5) at days -1 and +1 before and after each immunization. For dendritic cells tracking experiments, mice were immunized once with CTBA488 (40μg) and CT (1μg) and were sacrificed after 7 days.

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

EAE immunization:

The mice also received two 400 ng of intraperitoneal injections of pertussis toxin (List Biological Laboratories) in 100 µl of PBS immediately and 48 h later.

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)

Antibody variable region cloning and humanization:

The m1B7 and m11E6 variable region genes (17) were amplified from hybridoma cells by RT-PCR using degenerate primers and cloned into pAK100 as described (39). Positive clones were identified by monoclonal phage ELISA using a PTx-coated plate (1 mg/ml in PBS; List Labs), followed by sequencing.

ELISA and binding assays:

For indirect PTx ELISAs, a high-binding 96-well ELISA plate (Costar) was coated with 1 µg/ml PTx. The plate was blocked with milk for 1 hr, followed by incubation with duplicate anti-PTx antibody dilutions from 50 g/ml for 1 hr at 25°C. …

…PTx was injected in duplicate at 30 µL/min or 100 µL/min (m1B7, ch1B7), with concentrations between 5200nM diluted in running buffer (PBS or HBS, pH 7.4, 0.05% Tween). …

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)

Surgical procedures:

To visualize regenerating axons, cholera toxin subunit B (CTB; List Biological Laboratories) was injected using a protocol modified slightly from that in the previous report…After the sciatic nerve between the thigh muscles was exposed, a small incision was made on the perineurium just proximal to the trifurcation site. Four microliters of unconjugated CTB solution (0.1% in PBS) were slowly injected using the Hamilton syringe through the perineural incision and animals were killed 5 d after the injection.

p>Tissue processing and immunohistochemistry:

For immunohistochemistry, tissue sections were treated with 10% normal serum and 0.3% Triton X-100 for 1 h, and then primary antibodies dissolved in the same blocking solution were applied at 4°C overnight. The primary antibodies were…and goat anti-CTB (1:10,000; List Biological Laboratories).

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