Citations

Reagents:

LPS (catalog 421) was purchased from List Biological Laboratories (Campbell, CA). …

Cell culture, endotoxin assay, fluorescein diacetate assay, and cytokine ELISA:

Bone marrowderived macrophages (BMDM) were harvested from femurs of male C57BL/6. …On day 7, macrophages were left untreated (nontolerant) or stimulated with 5 endotoxin unit (EU)/ml LPS or 20 mg/ml LTA for 24 h (tolerant). At 24 h, cell cultures were washed twice with warm PBS and given fresh media or conditioned media with 5 EU/ml LPS (LPS/LPS incubation time or LTA/LPS-incubation time). After various times of incubation, media were tested for cytokine content. To ensure the same activity of LPS from different lots, we measured the concentration of LPS by Endosofe-PTS kinetic reader (Charles River Laboratories). 

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

Animal manipulations:

Wild type C57BL/6 mice used for most of these experiments…Each immunized mouse also received 300 ng of Bordetella pertussis toxin (List Biological Laboratories, Campbell, California, USA) injected intraperitoneally in 0.15 mL of phosphate buffered saline (PBS) on day 0 and day 2 post-immunization. Mice were examined daily for signs of EAE and disease severity was rated in each animal using a standard six-point scale as follows: …

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:

… tuberculosis H37Ra (Difco Laboratories). Animals were also injected intravenously (iv) on the day of, and two days following immunization, with 400 ng of Bordetella pertussis toxin (List Biologicals). …

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)

Injected tracers:

In most cases different fluorescent cholera toxin B-chain (CTB) tracers were used, either singly or in paired combinations in the same bird: …Biotinylated dextran amine (BDA, Invitrogen, Eugene, OR; 10K MW, 10% in PBS) was also used in some cases, in combination with a second injection of unconjugated CTB (List Biological Laboratories, Campbell, CA; 1% in PBS) at another site. BDA and CTB were injected either iontophoretically using a Stoelting high voltage current source (24 A, positive current) or via air pressure. …

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

Surgery and axonal tracing:

Five weeks after the surgery to generate dorsal column lesions was performed, animals were re-anesthetized with an XAK cocktail and an incision was made into the upper hindquarters to expose the sciatic nerve. A 10µL Hamilton syringe was inserted into each sciatic nerve at the junction of the fibial and tibial branches and 3 µL of cholera toxin B subunit (CTB;10 µg/L; List Biological Laboratories, Campbell, CA) was injected to label ascending sensory axonal projections. Animals were sacrificed three days after CTB injection.

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

Tracer injections:

Males and females MC4-R-GFP mice were deeply anesthetized with ketamine (5 mg/100 g) and xylazine (1 mg/100 g). The animals (n=10 males, n=4 females) received unilateral stereotaxic injections of the retrograde tracer cholera toxin b subunit (CTb 1%; List Biological Laboratories) into the PVH (0.43 mm from bregma; +0.23 lateral; 4.75 mm from surface of the skull). The retrograde tracer was injected with a glass micropipette and air pressure injection system as described previously …

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

… Injection site, gt. CTb, 1:50,000, List Biological Laboratories, Campell, CA, …

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

Experimental autoimmune encephalomyelitis (EAE)

Age- (5-8 weeks) and sex-matched mice were immunized subcutaneously with 100 g MOG₃₅₅₅ peptide (MEVGWYRSPFSRVVHLYRNGK) emulsified in CFA (Difco Laboratories) with 500 g Mycobacterium tuberculosis on day 0. Mice also received 200 ng pertussis toxin (List Biological Laboratories) by intraperitoneal injection on days 0 and 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)

ET Assays.

EF was obtained from List Biological Laboratories. J774A.1 cells in 384-well plates were incubated with 11 nM PA and 3.7 nM EF for 5 h at 37 C, and the concentration of cAMP was measured by cAMP enzyme immunoassay…

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).

Materials and Methods:

…Coronal sections of 40 m were cut on a freezing microtome, after which the sections were post-fixed in 4% PFA at 4C for 4 hours. Free-floating sections were then washed several times in 0.5% PBST (PBS tween), and then washed several times in 2.5% normal rabbit serum (diluted in 0.5% PBST). Sections were incubated in goat anti-CTB (#703, List Biological Laboratories, Campbell, CA) at 14000 in 2.5% normal rabbit serum diluted in 2% PBST for 96 hours at 4C. …

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