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Page 608 out of 618

Intranasal Delivery of Cholera Toxin Induces Th17-Dominated T-Cell Response to Bystander Antigens

Jee-Boong Lee, Ji-Eun Jang, Man Ki Song, Jun Chang
Product: Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Reagents:

CT and CTB were purchased from List Biological Laboratories (Campbell, CA). ...

Immunization:

For peptide immunization, 26106 OT-II splenocytes were i.v. injected into B6 mice, and then recipient mice were i.n. immunized with 40 mg of OVA peptide plus 30 mg of CpG ODN, 0.2 mg of CT, or 2 mg of CT or CTB in PBS. ...

Cytokine assays:

Dendritic cells (DCs) were generated from bone marrow of B6 or BALB/c mice by culturing in complete RPMI medium containing 10% FBS supplemented with 10 ng/ml recombinant GM-CSF and IL-4 (R&D, Minneapolis, MN). After 7 days of culture, non-adherent cells were harvested by gentle pipetting, and DCs were enriched by density gradient centrifugation over Percoll medium. After purification, DCs were further incubated with PBS or CT (100 ng/ml) for 3 days and the culture supernatants were harvested at the indicated time points. ...

 

Author did not specify which Cholera toxin was utilized.  List Labs Product #101B/C - Cholera Toxin from Vibrio Cholerae has been discontinued; however, List Labs still provides Product #100B - Cholera Toxin (AZIDE-FREE) from Vibrio cholerae.  

To follow, are our recommendations for transitioning from Product #101B/C to Product #100B:

Product #101B, Cholera Toxin from Vibrio cholerae (1 mg), has been discontinued.  Product #100B, Cholera Toxin (AZIDE-FREE) from Vibrio cholerae, may be substituted for Product #101B, if the user accounts for the formulation differences.  Both products contain the same quantity of toxin per vial (1.0 mg).  For Product #100B, reconstitution with 1.0 mL of water results in a 1 mg/mL toxin solution in a buffer with the following component concentrations:  0.05 M Tris, 0.2 M NaCl, 0.001 M Na2EDTA at pH 7.5.  For Product #101B, reconstitution with half that amount of water (0.5 mL) resulted in a 2 mg/mL toxin solution with the same concentrations of the buffer components, and in addition, Product #101B contained 0.003 M NaN3.  The user may add the sodium azide preservative to product #100B, if desired.

Product #101C, Cholera Toxin from Vibrio cholerae (2 mg), has been discontinued.  Product #100B, Cholera Toxin (AZIDE-FREE) from Vibrio cholerae, may be substituted for #101C, if the researcher accounts for the formulation differences.  While Product #101C contained 2.0 mg per vial, Product #100 contains 1.0 mg per vial.  When Product #100B is reconstituted with 1.0 mL of water, the resulting 1 mg/mL toxin solution will have buffer components with the following concentrations:  0.05 M Tris, 0.2 M NaCl, 0.001 M Na2EDTA at pH 7.5.  For Product #101C, reconstitution with less than half that amount of water (0.4 mL) resulted in a 5 mg/mL toxin solution in the same buffer, and in addition, Product #101C contained 0.003 M NaN3.  The user may add the sodium azide preservative to product #100B, if desired. 

Large quantities of Product #101B or Product #101C may be obtained through Bulk Requests and Customer Orders

 

Author did not specify which CTB was utilized.  List Labs has provided Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae) and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt).  The former has been discontinued.  The latter may be substituted for the former, if the researcher takes into account the difference in the buffer in which each product is provided and the amount of protein per vial.  When Product #103B was reconstituted with 200 μl of water, it contained 1 mg of protein in 0.05 M Tris, 0.2 M NaCl, 0.001 M Na2EDTA, 0.003 M NaN3, pH 7.5.  When Product #104 is reconstituted with 0.25 ml water, it contains 0.5 mg of Cholera Toxin B Subunit (choleragenoid) in 0.01 M Sodium Phosphate at pH 7.5.

PubMed ID: 19360100

Calcineurin Inactivation Leads to Decreased Responsiveness to LPS in Macrophages and Dendritic Cells and Protects Against LPS-Induced Toxicity In Vivo

Jennings C, Kusler B, Jones PP

Reagents:

... LPS lyophilized powder (List Biological Labs) was dissolved and diluted in phosphate buffered saline (PBS). As lethality varied among lots of LPS, when a lethal dose of LPS was desired for challenge, each lot of LPS was tested for LD50 before each experiment by injecting naïve mice with varying doses of LPS (5–10 mice per dose group) and assessing survival. The lowest dose that yielded 80–100% death was used for the LPS challenge dose in the subsequent experiment.

LPS challenge

All mice were observed every hour for the first 24–48 hours following the lethal LPS challenge. Mice were observed for characteristics of LPS toxicity (ocular exudates, diarrhea, increased respiratory rate) as well as moribund status (inability to sustain body temperature, ataxia or tremors, diaphragmatic breathing). In order to assess recovery, mice were given a score from 0–3 based on ambulation as an indication of overall health status: 0 = unable to walk, 1= take steps to walk, 2 = able to walk quickly but not run, and 3 = able to run/normal ambulation.

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

PubMed ID: 19318421

Efficient Neutralization of Antibody-Resistant Forms of Anthrax Toxin by a Soluble Receptor Decoy Inhibitor

Sharma S, Thomas D, Marlett J, Manchester M, Young JA
Product: Anthrax Protective Antigen (PA), Recombinant from B. anthracis

We next tested the ability of RDI to neutralize LeTx containing an antibody-resistant form of PA (PAL685A) in a rat model system. Male Harlan Sprague-Dawley rats (180 to 200 g; Harlan, Indianapolis, IN) were anesthetized with isofluoranes and inoculated with 500 μl LeTx mixture by intravenous injection in the tail vein. The anthrax LeTx was prepared for each group by mixing 12 μg LF (List Biological Laboratories, Campbell, CA) with 40 μg PA (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).

PubMed ID: 19075066

Ethanol Exposure Impairs LPS-Induced Pulmonary LIX Expression: Alveolar Epithelial Cell Dysfunction as a Consequence of Acute Intoxication

Walker JE Jr, Odden AR, Jeyaseelan S, Zhang P, Bagby GJ, Nelson S, Happel KI

Alcohol administration and intratracheal injection:

Mice received intraperitoneal (i.p.) injection with ethanol [4.0 g/kg; 20% v/v ethanol in sterile phosphate buffered saline (PBS)] or an equivalent volume of PBS. Thirty minutes after i.p. injection, animals were anesthetized with isoflurane for exposure and direct intratracheal (i.t.) administration of 10 μg Escherichia coli LPS (List Biological Laboratories; Campbell, CA). n ≥5 per group for all in vivo experiments.

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

PubMed ID: 19053978

Genetic Vaccines for Anthrax Based on Recombinant Adeno-associated Virus Vectors

Liu TH, Oscherwitz J, Schnepp B, Jacobs J, Yu F, Cease KB, Johnson PR
Product: Anthrax Protective Antigen (PA), Recombinant from B. anthracis

For the procurement of positive control hyperimmune anti-PA and anti-LF serum for use in the western blotting and as positive controls for the LeTx neutralization assays, rabbits were immunized with 250 µg of either soluble PA83 or LF in an emulsion with CFA (List Biological Laboratories, Campbell, CA). Rabbits were then boosted three times at 2–3-week intervals with 125 µg of the respective immunogens in an emulsion with incomplete Freund's adjuvant ...

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).

PubMed ID: 19002162