Citations

Reagents and Instruments:

…LPS from E. coli O111:B4 was obtained from LIST Biological Laboratories. …

LPS stimulation, Cytokine Production, and NF-B Activation:

Concentrations of secreted IL-8 in the supernatants after LPS stimulation were measured by ELISA. Briefly, 310⁵ cells of each cell line were seeded in 24 well plates in complete media containing 100 g/ml Zeocin. After 1620 hours incubation, cells were stimulated with 0, 0.1, 0.5, 2 and 20 ng/L LPS for 24 hours. Concentrations of human IL-8 were measured by Human CXCL8/IL-8 DuoSet ELISA kit. Each experiment was repeated three times.

Intracellular concentrations of human IL-8, phosphorylated and total NF-B p65 after LPS stimulation were also determined by intracellular flow cytometry analysis. Briefly, 1620 hours before stimulation, 1.010⁶ cells of each cell line were seeded in 6-well plates in complete media containing 100 g/ml Zeocin. …

• Product #201 – LPS from Escherichia coli O111:B4

Macrophage isolation, culture and transfection:

Cells were transfected with poly(dA:dT) (2 ng/mL; Sigma), MDP (20 ug/mL; Invivogen), Salmonella thyphimurium flagellin (140 ng/mL; Invivogen) and Bacillus anthracis Lethal toxin and protective antigen (1 ug/mL; List Biological Laboratories).

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

Cell lines, bacterial strains, toxin, and antibodies:

…Purified C. difficile toxin B was purchased from List Biological Laboratories (Campbell, CA). …

RTCA version 2 assay:

…After the CI reached a plateau, the CI for each sample well was normalized to one at the last measurement time point prior to toxin treatment and inoculated with eluted samples or purified CDT to be tested. The nCI was then automatically monitored at 5-min intervals for 36 h… Each run included one positive control with 100 ng/ml purified CDT in duplicate and one negative control in quadruplicate.

The time point at which the nCI dropped by 30% was considered a positive step time (PST). PST was used to calculate the toxin concentrations in samples by comparing standard curves which were developed by monitoring the nCI at serial 3-fold dilutions of purified CDT ranging from 0.01 to 787.32 ng/ml and plotting the PST versus the purified CDT concentration. The limit of detection of the RTCA version 2 assay was determined by testing 11 3-fold serial dilutions of purified CDT (concentrations ranging from 0.01 to 787.32 ng/ml). All testing was done in triplicate.

• Product #155 – Toxin B from Clostridium difficile

Intravitreal Injections and Factors:

Balb/c mice were anesthetized as described, and the vitreous of the right eye (treated) was injected with Rho GTPases activator or inactivators and/or growth factors (GFs), …Other factors injected included the general inhibitor for Rho GTPase, Toxin A, from C. difficile (10 ng/eye, No. 152C; List Biological Laboratories, Inc., Campbell, CA, USA), …

• Product #152C – Toxin A from Clostridium difficile

Iso- lated epithelial cells were suspended in a keratinocyte culture medium (KCM) composed of a basal mixture of 3 parts Dulbeccos modified Eagles medium (DMEM) and …1 nM cholera toxin (List Biological Laboratories, Campbell, CA), …

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

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

Reagents:

…LPS from E. coli serotype 0111:B4 was purchased from List Biological Laboratories (Campbell, CA). …

Animal protocols:

C57BL/6 and TLR-4^/ mice (78 weeks age) …Using methods that were previously described, the intratracheal (IT) instillation of live E. coli, E. coli 0111:B4 LPS (1 g/g), or PBS was performed in mice anesthetized with 34% isoflurane …

• Product #201 – LPS from Escherichia coli O111:B4

EAE:

…Mycobacterium tuberculosis H37Ra (Difco). Each mouse was subsequently administered 500 ng pertussis toxin (List Biological Laboratories) ip on days 0 and 2 after MOG 3555 peptide immunization. The EAE clinical manifestations…

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:

… Pertussis toxin (400 ng) (List Biological Laboratories, Campbell, CA, USA) in PBS and 50 mM NaCl was administered ip on the day of immunization and again after 24 h. For the treatment of EAE, EB (25 and 50 mg/kg) or PBS (vehicle control) was administered po daily started …

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 and evaluation of EAE:

Mice were immunized for the induction of EAE using either a 1 or 2 immunization protocol. …On the day of immunization, each mouse received 200 ng of PTX (List Biological Laboratories, Campbell, CA, USA) by intervenous 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)