Citations

EAE:

… 100 µg pure MOG 35-55 in complete Freund’s adjuvant (CFA, 8 mg/mL Mycobacterium tuberculosis, M. tb) (IFA from Thermo Fisher and M.tb from BD, Sparks, MD) was injected into the abdomen subcutaneously on day 0. Pertussis toxin (List Laboratories, Campbell 

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 induction and ERβ-ligand treatment

Female mice were immunized subcutaneously with MOG, amino acids 35–55 (200 μg per mouse, Mimotopes) emulsified in Complete Freund’s Adjuvant, supplemented with Mycobacterium tuberculosis H37Ra (200 μg per mouse, Difco Laboratories), over two sites drained by left inguinal and auxiliary lymph nodes in a total volume of 0.1 ml per mouse (Day 0). Pertussis toxin (500 ng per mouse, List Biological Laboratories) was injected intraperitoneally on Day 0 and Day 2. On Day 7, a booster immunization was delivered over contralateral lymph nodes. Ages of female mice for EAE induction were 8 to 12 weeks.

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)

Chemicals, reagents, equipment and safety:

… Recombinant LF (rLF) was purchased from List Biological Laboratories (Campbell, CA).

Sample preparation:

The MS method for anthrax LF relies on three simple steps that each add a layer of sensitivity and specificity (Fig. 1). LF-MABs selectively purify and concentrate total-LF (LF + LTx) (Step 1). Immobilized LF is reacted with an optimized peptide substrate which it hydrolyzes producing two unique LF-specific product peptides (Step 2). Because each LF molecule can continuously hydrolyze peptide substrate, this step allows amplification of the resulting signal and improves detection. Analysis of the cleaved peptides by high-sensitivity/high-resolution isotope-dilution (ID) MALDI-TOF MS (Step 3). This strategy has been successfully applied to several enzymatic protein toxins [22].

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).

Cell culture and treatment:

… For stimulation, cells were exposed to 100 ng/ml LPS (ultrapure smooth LPS from E. coli O111:B4,
List Biological Laboratories, Campbell, CA).

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

Localized EAE Mouse Model:

Adult female C57BL/6J mice (68-weeks-old) were subcutaneously immunized with an emulsion of 50 g myelin oligodendrocyte glycoprotein (MOG)3555 … All mice received an intravenous injection of 200 ng pertussis toxin (List Biological Laboratories) in 100 µL PBS at 48 h after injection of TNF- and IFN-. 

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)

EAE induction. Mice were s.c. injected into both flanks with a total of 200 μl of emulsion containing either 300 μg of MOG35–55 (Synpeptide) or 500 μg of bMOG (see above) dissolved in saline and mixed with an equal volume of complete Freund’s adjuvant containing 500 μg of killed Mycobacterium tuberculosis H37 RA (Difco Laboratories). Mice were also i.p. injected with 20 μg/kg of PTX (List Biological Laboratories) immediately and 2 days after 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)

Diphtheria Toxin List Biological Labs 150

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

Mouse primary corneal epithelial cells (MPCEC)

… Each cornea was cut in half and placed in a 35 mm dish (Fisher Scientific, PA) with the epithelial side up. After approximately 5 minutes in the laminar air-flow culture hood, the half corneal button attached itself to the bottom of the plate, and 1.5 mL of DMEM (Life technology, NY) with 10% serum containing 40 μg/mL gentamicin (Life technologies, NY), 1% ITS (BD Biosciences, Bedford, MA), and 100 ng/mL cholera toxin (LIST Biological Laboratories, Inc., Campbell, CA) were added and the tissue was placed in a humidified incubator at 37 °C with 5% CO2. Culture medium was replaced every 2 days. After 7 to 10 days cells were nearly 100% confluent. …

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

EAE:

Mycobacterium tuberculosis (H37Ra; Difco Laboratories). In addition, all mice were given 200 ng pertussis toxin (List Biologicals Laboratories) intraperitoneally on days 0 and 2 post-immunization (pi). Mice were graded for clinical 

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)

Antigens:

… Recombinant protective antigen (rPA) was obtained from List Biological Laboratories (Campbell, CA). Each vial was reconstituted  

Quantification of anti-PA IgG antibodies:

Wells of an Immulon 2 HB, 96-well round bottom microtiter plate (Southern Biological, Birmingham, AL) were coated with 100 l of a coating solution (rPA diluted at 1 g/ml in a 0.01 M PBS solution, pH 7.4). …

Titration Curve:

Eight rPA-coated microtiter plates were prepared. Following overnight incubation, approximately 200 l of REF plasma that had been diluted to 1:500 was added to the first well of three rows on each plate then two-fold serial dilutions carried out. …

Specificity:

In the second experiment, two rPA coated microtiter plates were prepared, except that ovalbumin (100 l diluted at 1 g/ml in a 0.01 M PBS solution), a heterologous protein to B. anthracis PA, was also used to coat six wells in each plate. …

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