Bacterial Toxin Research Citations

Anthrax lethal toxin neutralization assay:

Samples collected after release were characterized for toxin-binding activity via an anthrax LeTx neutralization assay that was performed using decreasing amounts of the released samples. Specifically, the biological activity of PANG was measured using a LeTx neutralization assay as described previously with modifications.³⁵ J774A.1 (ATCC TIB-67, Manassas, VA) cells were plated in 96-well flat-bottomed tissue culture plates at 2.5 10⁴ cells/well in 50 L and incubated for 1619 h at 37C with 5% CO₂. Test samples were diluted and titrated on the plated cells in the presence or absence of LeTx (List Biological Laboratories, Campbell, CA). …

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).

Drug administration:

…In other cases they were treated with both flumazenil (10 M) and/or botulinum toxin B (5 nM) across a time-course (1, 2, 4, 6 or 24 h). In all cases, cells were harvested at the end of the treatment so that all groups would be processed in parallel (Inset, Fig. 2). Other drugs used included …and botulinum toxin B was from List Biological Laboratories, Inc. (Campbell, CA).

Reagents and Materials:

All reagents purchased were of the highest grade possible. … The BoNT-A holotoxin was purchased from List Biological Laboratories Inc., Campbell, CA, USA. …

BoNT-A Sera Spike and Dilution Series:

A BoNT-A holotoxin stock was diluted to 1 g/mL in saline to make a working solution. A pool of sera representing each animal species was spiked initially to 100 ngmL¹ from the BoNT-A working solution and serial dilutions using the corresponding animal sera were used to make the dilutions used in the assay titration.

Mice and immunizations:

Adult mice (aged between 7 and 13 weeks) were immunized…and 10 µg CT (List Biological, Campbell, CA) in 200l phosphate-buffered saline (PBS) using a feeding needle that was placed through the mouth into the stomach.

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

Immunofluorescence labeling:

…We used similar double-immunofluorescence protocols to label CTb with FoxP2 or c-Fos. In a 1:3 series, CTb was labeled using a goat polyclonal antiserum (1:5,000; List Biological Laboratories, no. 703, lot no. 115037) in combination with rabbit anti-c-Fos. …

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

Reagents:

Wild-type LPS from E. coli K12 was obtained from List Biological Laboratories (Campbell, Calif). …

Results:

Wild-type LPS stimulated CD14-independent TNF- and IL-10 secretion

We next set out to investigate whether alteration of the O-Antigen affects interactions between CD14 and the LPS molecule. TNF- and IL-10 secretion was measured in PBMCs pretreated with blocking antibody to CD14 and stimulated with 100 ng/mL LPS, RF5, or DPL. CD14 blockade resulted in significant inhibition of TNF- and IL-10 secretion from cells stimulated with 100 ng/mL wild-type LPS, as has been previously demonstrated ⁽¹⁴⁾, but modest amounts of cytokine secretion did occur. With attenuation or removal of the O-Antigen, secretion of both cytokines in the presence of CD14 antibody decreased to control levels (Fig. 5). DPL, even in 1000 ng/mL concentrations, was unable to stimulate secretion of TNF- in the presence of CD14 blockade (data not shown).

• Product #314 – LPS from Escherichia coli K12 strain LCD25

… After 30 min, monocytes were treated with 0.1 ng/ml LPS (S. minnesota R595, List Biological Laboratories, Campbell, CA, USA)

• Product #304 – LPS from Salmonella minnesota R595 (Re)

… We used smooth-type LPS from E. coli O55: B5 (List Biological Laboratories, CA), Rc-type LPS from E. coli J5 (List Biological Laboratories), …

• Product #203 – LPS from Escherichia coli O55:B5
• Product #301 – LPS from Escherichia coli J5 (Rc)

Materials:

…Lipopolysaccharide (LPS) was obtained from List Biological Laboratories (USA). …

Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza (USA) at passage 1 and maintained in EBM2 Bullet kit growth medium at 37C in a humidified atmosphere of 5% CO2. In all experiments, cells were used at passage 4~9. HUVECs were plated at 90~95% confluence treated for 1 h with 1, 10, and 20 M of quercetin prior to LPS (1 g/mL) stimulation for 1 or 8 h. …

Author did not specify which List Labs LPS product was utilized in their research.  List Labs provides the following LPS products:  https://listlabs.com/product-information/lipopolysaccharides/

… For activation of monocytes, 200-L PBMCs were incubated with 10 g/L ultrapure lipopolysaccharide (LPS) (List Biological Laboratories, Inc, Campbell, CA, USA) in separate 96-well plates. …

Author did not specify which List Labs LPS product was utilized in their research.  List Labs provides the following LPS products:  https://listlabs.com/product-information/lipopolysaccharides/