Bacterial Toxin Research Citations

… Anthrax lethal factor and edema factor were from 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)

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).

Reagents:

Recombinant expression and purification of LFn-FlaA was performed as previously described [19]. B. anthracis protective antigen (PA) and lethal factor (LF) were acquired from List Biologicals. …

Macrophage differentiation and stimulation:

…  For NLRP1b inflammasome activation, cells were stimulated with LeTx (1g/ml PA combined with 0.5 or 1g/ml LF). …

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 and Methods

Anthrax lethal factor (Product #172), and the chicken IgY polyclonal anti-LF antibody (Product # 769A) are
products of List Biological Laboratories, Inc. The Nunc-Immuno Tubes, Maxisorp (cat# 444202) used for LF
antibody coating and the dimethyl sulfoxide (DMSO) (cat # TS-20684) were purchased from ThermoScientific.
The 96-well, black, flat bottom, non binding plates used for the fluorescent plate assay were from Corning
(cat # 3991). Bovine plasma (cat # 7310806) was purchased from Lampire Biological Laboratories.

Sample Preparation: Stock solutions of the fluorogenic substrates, MAPKKide® Plus (± biotin), were made
2.5 mM in DMSO based on the peptide content determined by elemental analysis. The substrates were diluted
in assay buffer: 20 mM HEPES, pH 8.0 containing 0.1% Tween-20. The LF was added to neat bovine plasma.

LF Activity Assays:
Antibody Capture: The Nunc-Immuno Maxisorp Tubes were coated with 1 mL of a 10 μg/ml solution of
a chicken affinity purified polyclonal IgY antibody to anthrax lethal factor (List Prod # 769A).
The immuno-tubes were incubated with the IgY overnight at 2-8°C. After 5 washes with 1.5 mL each of PBS containing 0.05%
TWEEN-20 (PBST), the anti-LF coated tubes were exposed to 1 mL of a series of LF concentrations in neat
plasma. The tubes were incubated at 22°C for 1 hour. Tubes were then washed 3 times with 1.5 ml PBST and
1.0 ml of 10 μM MAPKKide® Plus (± biotin) was added. The reaction was allowed to proceed for 3, 5 and
22 hours at 37°C. At each time point 250 μl of the reaction mixture was removed from triplicate tubes and placed
in a 96 -well black plate. In the case of the biotinylated MAPKKide® Plus a 0.5 ml aliquot of each triplicate
was added to streptavidin-coated beads after 22 hours of exposure to LF. The mixture was gently rotated at
ambient room temperature for 1 hour and then read in the platereader.

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #769B – Anti-Lethal Factor from B. anthracis (Chicken lgY)
• Product #532 – MAPKKide® Plus (AMC) Specific Substrate for Anthrax Lethal Factor

… MAPKKide was obtained from List Biological Laboratories (Campbell, CA 

Author did not specify which List Labs MAPKKide was utilized; the following are available:

• Product #530 – MAPKKide Peptide Substrate (o-Abz/Dnp) for Anthrax Lethal Factor
• Product #531 – MAPKKide Peptide Substrate (DABCYL/FITC) for Anthrax Lethal Factor
• Product #532 – MAPKKide Plus (AMC) Specific Substrate for Anthrax Lethal Factor

Detection of Toxin Agents:

… As illustrated in Figure 4a, the derived GT1b LSPR chip utilized Au nanoparticles of 40 nm in size. In the present study, a heavy chain binding domain (BTX/A/Hc) was used instead of the neurotoxins (BTX/A) composed of Hc and Lc with potent toxicity.  To assess the LSPR biosensor unit, we examined the potential utility of the biosensor using different concentrations of RCA120, ricin (RCA60) and BTX/A/Hc in solution.

• Product #612A – Botulinum Neurotoxin Type A Heavy Chain, Recombinant, Binding Domain

… Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native, List Biological Laboratories, Cat#169A. Anthrax Protective Antigen (PA), Recombinant from B. anthracis, List Biological Laboratories, Cat#171E. …

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

List Labs Product #169A (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence; Lyophilized) has been discontinued; however, Product #169L (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence; Liquid) is available for purchase.

• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Briefly, heat-extracted CFA/I, 389 CS1, CS2, CS3, CS4 or CS5 fimbriae (200 ng per well) and LT (List Biological Laboratories, Inc., Campbell, CA; 100 ng per well) were coated to wells of 2HB plates …

Lethal Toxin (LT) consists of LF + PA:

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

Quantification of anti-PA IgG antibodies:

A 96-well plate (Immulon 2 HB Southern Biological, Birmingham, AL, USA) was coated with 100 l of rPA coating solution (1 g/ml in PBS, List Biological Laboratories, Inc. Campbell, CA, USA). …

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

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).

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