Bacterial Toxin Research Citations

… Mice were sensitized by intragastric administration of antigen plus cholera toxin (CT) as adjuvant once a week for six weeks. Feedings consisted of 1 mg of ovalbumin (OVA, grade V, Sigma Chemicals, St. Louis, MO) or ovomucoid (OM, Sigma) plus 10 µg CT (List Biologicals, Campbell, CA, USA) in 0.4 ml of 0.2 M bicarbonate. …

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

EAO induction:

Six-12 week old mice were immunized as previously described [9] with 10 mg of TH plus CFA (Sigma-Aldrich, St. Louis, MO) supplemented with 200 µg of Mycobacterium tuberculosis H37Ra (Difco Laboratories, Detroit, MI) in conjunction with PTX (List Biological Laboratories Inc., Campbell, CA). EAO was evaluated at 25-30 days post-injection. …

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)

Experimental Autoimmune Encephalomyelitis (EAE):

… Briefly, eight-week-old female SJL/J mice were anesthetized with ketamine/xylazine and were injected subcutaneously at two sites on the back with 0.1 mL solution containing 0.5 mg/mL proteolipid protein peptide 139151 (GenScript, Piscataway, NJ) emulsified in complete Freund adjuvant (CFA; Difco, Detroit, MI) containing 2.5 mg/mL Mycobacterium tuberculosis (Difco). Control mice were injected with equal volumes of PBS and CFA. All mice were injected with 200 ng pertussis toxin (List Biological, Campbell, CA) in 0.1 mL PBS intraperitoneally on day 0 (day of immunization) and again 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)

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

… Recombinant PA, EF, LF toxins and goat anti-PA primary antibody (Ab) were obtained from the List Biological Laboratories, Inc. … 

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

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Product #178A – Anthrax Edema Factor (EF), Recombinant from B. anthracis
• Product #173B – Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis

… In a typical experiment, cells before stimulation were washed with serum-free medium, incubated for 1 h, and then stimulated in the conditions indicated in figure legends. LT, ET, protective antigen, lethal factor, and edema factor were purchased from List Biologicals. …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Author did not specify which List Labs Anthrax Edema Factor was utilized.  List Labs provides Product #178A(Anthrax Edema Factor (EF), Recombinant from B. anthracis) and Product #173B (Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis).

PA-specific IgG ELISA:

Sera from individual rabbits were assayed in triplicate for the presence of PA-specific IgG by ELISA. Briefly, 96-well flat-bottomed Nunc Maxisorp plates (VWR International, Inc.) were coated with 0.1 g/well of PA (List Biological Laboratories, Inc.) overnight, blocked with 0.5% casein in phosphate-buffered saline (PBS) for 2 h at 37C, washed, and incubated for 1 h at 37C with the test serum. …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Materials and Methods – Anthrax Lethal Toxin:

Recombinant anthrax toxin PA and LF were obtained from List Biological Laboratories (Campbell, CA, USA). Antigens were snap frozen in liquid nitrogen and stored at 70 C in 10 L aliquots at a concentration of 1 mg/mL.

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Materials:

Anthrax rPA and recombinant LF rLF were obtained from List Biological Laboratories, Inc. (Campbell, California). …

Macrophage Toxicity Assay

Mouse macrophages are sensitive to LeTx(the combination of PA and LF )-induced necrosis.. Here, rLF was added to the reconstituted powder formulation and the activity/stability of rPA was evaluated by measuring the PA dependent toxicity of rLF using mouse macrophage J774A.1 cells …

Anthrax lethal toxin neutralization assay:

Mouse macrophages are sensitive to LeTx(the combination of PA and LF )-induced necrosis. An in vitro lethal toxin (LeTx) neutralization assay using J774A.1 mouse macrophage cells was performed to determine the functional titer of the anti-PA antibody present in rabbit serum. The details of the assay were reported previously ³⁵. Briefly, rabbit sera were incubated with PA for 1 hr at 37C to allow neutralization to occur, followed by incubation with added LF at room temperature for 1 hr. The serum-PA/LF mixture was then added to the plate containing macrophage cells. The starting serum dilution on the final plate is 1:128 or 1:2⁷ with a final concentration of 0.1875 g of LeTx (rPA+rLF) per ml. Percent neutralization was calculated based on (sample OD value LeTx standard OD value)/(cells only OD value-LeTx standard OD value) 100. The percent neutralization was plotted vs serum log₂ dilutions and fitted with a second order polynomial curve. The serum dilution able to neutralize 50% of LeTx (NT50) was calculated. …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Immunization and infection of mice:

Mice were immunized by subcutaneous (s.c.) injection with 10 g of recombinant PA (List Biological Laboratories Inc.) and 0.3% aluminum hydroxide gel (Sigma-Aldrich) in 100 l of vaccine 20 days before infection, with a booster given 10 days before infection. …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis