Citations

Bacterial Toxin Research Citations

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Page 19 out of 35
344 citations found

Evaluation of early immune response-survival relationship in cynomolgus macaques after Anthrax Vaccine Adsorbed vaccination and Bacillus anthracis spore challenge

Sivko, GS;Stark, GV;Tordoff, KP;Taylor, KL;Glaze, E;VanRaden, M;Schiffer, JM;Hewitt, JA;Quinn, CP;Nuzum, EO;

Product: Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Product: Anthrax Lethal Factor (LF), Recombinant from B. anthracis

Product: Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Product: LPS from Escherichia coli O111:B4

  • Materials and Reagents:

    Anthrax lethal toxins (PA and LF) and Escherichia coli O111:B4 LPS were purchased from the List Biological Laboratories.

    Author did not specify which List Labs Lethal Factor was utilized.  List Labs offers Lethal Factor Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

    RESULTS:

    HDAC8 inhibition restores the production of pro-IL-1 in response to LPS in LeTx-exposed murine macrophages. LeTx suppresses expression of various inflammatory cytokines through inactivating MEKs (3-5). Consistent with these reports, a sub-lethal dose of LeTx (100 ng/ml PA & LF, a.a. for 4 h) completely cleaved MEK1 and inhibited expression of pro-IL-1 induced by LPS (100 ng/ml) for more than 2 days in RAW264.7 macrophages (Fig. 1A). …

The MUC1 mucin specifically inhibits activation of the NLRP3 inflammasome

Ng, GZ;Sutton, P;

Product: Anthrax Lethal Factor (LF), Recombinant from B. anthracis

A Comparison of the Adaptive Immune Response between Recovered Anthrax Patients and Individuals Receiving Three Different Anthrax Vaccines

Laws, TR;Kuchuloria, T;Chitadze, N;Little, SF;Webster, WM;Debes, AK;Saginadze, S;Tsertsvadze, N;Chubinidze, M;Rivard, RG;Tsanava, S;Dyson, EH;Simpson, AJ;Hepburn, MJ;Trapaidze, N;

Product: Anthrax Edema Factor (EF), Recombinant from B. anthracis

In vivo dynamics of active edema and lethal factors during anthrax

Rougeaux, C;Becher, F;Ezan, E;Tournier, JN;Goossens, PL;

Product: Anthrax Lethal Factor (LF), Recombinant from B. anthracis

Soluble Expression and Characterization of Biologically Active Bacillus anthracis Protective Antigen in Escherichia coli

Suryanarayana, N;Vanlalhmuaka, ;Mankere, B;Verma, M;Thavachelvam, K;Tuteja, U;

Product: Anthrax Protective Antigen (PA), Recombinant from B. anthracis

  • Bacterial Strains and Reagents:

    … Activated protective antigen was procured form List Biological Laboratories (USA). …

    2.6. Binding to LF and Native-PAGE

    For the confirmation of functional activity of the expressed PA, 2g of trypsin digested PA was incubated with 2g of LF at room temperature for 30 minutes. The samples were run on 4% native polyacrylamide gel at 4C and visualized by staining the gel with coomassie brilliant blue.

    2.7. Animal Immunization

    Six female Balb/c mice were procured from Institutional Animal Care Center, DRDE, and were provided with food and water ad libitum. Each mouse was immunized subcutaneously with 10g PA adsorbed on aluminium hydroxide adjuvant on days 0, 14, and 28. Mice receiving only PBS were taken as control. All mice were bled prior to first immunization on day 0 and on 35th day. Serum was separated and stored at 20C until further use.

    2.8. Enzyme Linked Immunosorbent Assay

    To test the reactivity of PA immunized sera, indirect ELISA was performed by coating 1gmL1 PA in ELISA plate overnight at 4C. After blocking with 5% skim milk for one hour, serial twofold dilution of PA immunized sera starting from 1:1000 was added and incubated for one hour at room temperature. …

    2.9. Cell Culture

    Mouse macrophage cell line J774.1 was procured from National centre for cell sciences, Pune, India and was maintained in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% fetal bovine serum (FBS) and 100UmL1 penicillin and streptomycin as antibiotics. One day before the experiment 4 104 cells were seeded in a 96 well plate and incubated at 37C in presence of 5% humidified CO2. The next day spent media was replaced by a fresh medium containing gradient concentration of soluble PA and native PA of B. anthracis from List Biologicals (0.005gmL1 to 5gmL1) and was incubated at 37C along with 0.125gmL1 LF. After 4 hours, 20L of 5mgmL1 MTT (3-(4,5 dimethylthiazol-2 yl)-2,5 diphenyl tetrazolium bromide) was added to each well and further incubated for 30 minutes.

Substrate For Specific and Quantitative Detection of Anthrax Lethal Factor in Plasma

Shine, N., Suryadi, K.

Product: MAPKKide® Plus (AMC) Specific Substrate for Anthrax Lethal Factor

  • Materials and Methods

    Anthrax lethal factor (Product #169), and the chicken IgY polyclonal anti-LF antibody (Product # 769A) are products of List Biological Laboratories,Inc.

    Sample Preparation:
    Stock solutions of the fluorogenic substrate, MAPKKide® Plus, were 1.25 mM in DMSO based on the peptide content
    determined by elemental analysis. The substrate was diluted in assay buffer: 20 mM HEPES, pH 8.0 containing 0.1% Tween-20. For the microplate
    assays, the LF was dissolved in neat bovine plasma and diluted 1:10 in assay buffer. For the HPLC assays, the LF was added to neat bovine plasma and not diluted.

    LF Activity Assays:
    Microplate reader: Assays were performed on a SPECTRAmax GEMINI XS fluorescence microplate reader (Molecular Devices). The
    cleavage reaction was initiated by addition of the substrate, MAPKKide® Plus. The concentration was optimized to minimize background fluorescence while maintaining measureable cleavage. For all experiments the time-dependent increase in fluorescence was monitored at 37°Chourly for 5 or 6 hours followed by an additional 18 to 18.5 hr overnight incubation at ambient temperature. The excitation wavelength was set to 368 nm and emission to 452 nm with a cutoff filter at 435 nm.

    HPLC:
    The C8 Starwell Maxi Nunc-Immuno Module Plates were coated with 150 μl of a 10 μg/ml solution of a chicken affinity purified
    polyclonal IgY antibody to anthrax lethal factor (List Prod # 769A)
    . Plates were incubated with the IgY overnight at 2-8°C and washed three times with 0.1M Glycine-HCl, pH 2.5. This wash was included to liberate residual LF retained after the affinity purification of the antibody and was necessary to minimize the background. After 6 washes with PBS containing 0.05% TWEEN-20 (PBST), the anti-LF coated wells were exposed to 300 μl of a series of LF concentrations in neat plasma. The plates were incubated at 22°C for 2 hours. Plates were then washed 6 times with PBST and 250 μl of 1.25 μM MAPKKide® Plus was added. The reaction was allowed to proceed for 2, 3.5, and 5 hours at 37°C and overnight at ambient temperature. At each time point 200 μl of the reaction mixture was removed from replicate wells and placed in HPLC sample vials. HPLC was performed using a Zorbax Eclipse Plus C18 reverse phase column, 4.6 x 150 mm (Agilent) and a guard column containing the same resin in a Varian ProStar HPLC system (Agilent). Solvent A was 0.1% TFA in water and solvent B was 0.1% TFA in acetonitrile. The 16 minute HPLC method was as follows: 25% B for 0.75 minutes; 25 to 45% B in 4.75 minutes; 45 to 100% B in 0.75 minutes; 100% B for 3.75 minutes; 100 to 25%B in 0.67 minutes and 5.34 minute equilibration with 25% B. The column effluent was monitored using an Hitachi fluorescence detector with excitation set to 350 nm and emission at 450 nm to detect the free coumarin fluorophore cleaved from MAPKKide® Plus. The injection volume was 20 µl. The 7-amido-4-methylcoumarin (AMC) peak retention time was 4.8 minutes.

    Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence
    Product #769B – Anti-Lethal Factor from B. anthracis (Chicken lgY)
    Product #532 – MAPKKide® Plus (AMC) Specific Substrate for Anthrax Lethal Factor

Product: Botulinum Neurotoxin Type B Light Chain, Recombinant