Bacterial Toxin Research Citations

 Materials:

The dominant negative inhibitor antigen was produced by Pharmaceutical Solutions LLC
(Bloomington, IN) as a lyophilized powder in sealed glass vials. The vials of the DNI protein
contained 25 mg of protein, 113 mg mannitol, 33 mg sucrose, and 2.4 mg dibasic sodium
phosphate. They were reconstituted with 3.3 ml of water for injection (WFI), the
reconstituted protein concentration was 91 M . 10 mg of recombinant protective antigen was
also purchased from List Biological Laboratories Inc (Campbell, CA). …

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

EAE immunization and clinical evaluation:

Age-matched WT and Tph-1/mice were immunized subcutaneously with 125 µg MOG3555 peptide (Peptides International) emulsified in IFA (Sigma-Aldrich) supplemented with 0.5 mg/mouse Mycobacterium tuberculosis H37 (Difco Laboratories) on day 0 and an i.p. injection of 200 ng pertussis toxin (List Biologicals) on days 0 and 2. …

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)

Experimental Procedures 

Reagents Ultrapure LPS (Escherichia coli 0111:B4) was from List Biological Laboratories, Inc. (Vandell Way, CA) …

Animals:

… In this study, we further elucidate the pathways involved in NO-mediated TACE activation and show that LPS stimulation upregulates iNOS expression in mouse hepatocytes, which leads to TACE and iRhom2 phosphorylation in a cGMP/PKG dependent manner. …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

… Quantitative RT-PCR. Peritoneal Ms were treated with or without 10 ng/ml of ultrapure E. coli LPS (0111:B4, List Biological Laboratories) for 2 h. …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

Induction of EAE in Mice by Immunization with MOG and Adjuvant and Treatment with HspB5 :

EAE was induced by procedures previously described (35). Briefly, EAE was induced in female C57BL/6J mice (Jackson Laboratories, Sacramento, CA) at 9 weeks of age by subcutaneous immunization in the flank with an emulsion containing 200 µg of myelin oligodendrocyte glycoprotein(3555) (MOG3555; MEVGWYRSPFSRVVHLYRNGK) in saline and an equal volume of complete Freund’s adjuvant containing 4 µg/ml of mycobacterium tuberculosis H37RA (Difco Laboratories, Detroit, MI). All mice were administered 400 ng of pertussis toxin (List Biological, Campbell, CA) intraperitoneally at 0 and 48 h post-immunization. …

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)

Los ratones fueron sensibilizados va oral utilizando agujas intragstricas (01-2902B; Fisher Scientific) y una solucin de 1 mg de protenas de man ms 10 µg de la toxina clera (CT; List Biological Laboratories) disueltos en 10 µl de agua estril. …

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

Immunohistochemistry for tracers:

… For double labeling with CTb, following BDA detection, free-floating sections were blocked 1 hr in 0.3% PBST with NRS (VECTASTAIN Elite Kit, Vector Labs). Blocked sections were incubated 2 hrs at RT in goat anti-CTb (110000 dilution, List Biological Laboratories) followed by 1 hr in rabbit anti-goat biotinylated secondary antibody (VECTASTAIN Elite Kit) and then reacted 30 min in ABC solution. DAB staining was as above, but for 3 min without nickel to give a brown reaction product. …

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

Tissue Processing, Analysis, and Immunohistochemistry

Stellate ganglia:

… Tissue sections were washed in 10 mM Tris, 0.9% NaCl, 0.05% thimerosal in 10 mM phosphate buffer (TPBS), pH 7.4, containing 0.3% Triton X-100 for 3 10 min then incubated in 10% heat-inactivated normal horse serum (NHS, Invitrogen) in TPBS-Triton for at least 1 h. The sections were then incubated in goat anti-CTB antiserum (1:25000, List Biologicals) in TPBS-Triton containing 10% NHS for 3 days at room temperature. …

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

Materials:

… Nicked anthrax PA 63 from B. anthracis was obtained from List Biological Laboratories Inc., Campbell, CA. …

Binding Experiments:

The binding of the His-tagged proteins to the C2II-channel and the binding component PA₆₃ was investigated with titration experiments similar to those performed previously to study the binding of 4-aminoquinolones to the C2II- and PA₆₃-channels and EF and LF to the PA₆₃-channel in single- or multi-channel experiments [38][40]. The aqueous phase contained always 150 mM KCl, buffered with 10 mM MES-KOH to pH 5.5 to pH 6. The C2II- and PA₆₃-channels were reconstituted into lipid bilayers. About 60 minutes after the addition of either activated C2II or PA₆₃ to the cis-side of the membrane, the rate of channel insertion in the membranes was very small. Then concentrated solutions of His-tagged proteins were added to the cis-side of the membranes while stirring to allow equilibration.

• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Intra-ocular injections:

The B subunit of the Cholera Toxin complex (CTB) has been shown to be a highly sensitive anterograde tracer for RGCs (Angelucci et al., 1996; Reiner et al., 1996; Matteau et al., 2003), and therefore the preferred tracer for this study. Rats were first anesthetized with 25% isoflurane mixed with oxygen at a flow rate of 1l/min., using an isoflurane vaporizer (Smiths Medical, Dublin, OH, USA) While maintained at the appropriate level of anesthesia, subjects were subcutaneously injected with buprenorphine (0.06mg/kg rat weight). Subjects then received, via syringe, 56 2l injections of either unconjugated CTB in one eye, or fluorophor-conjugated CTB in both eyes.

The monocular injections were administered into the vitreous chamber of the left eye only, and comprised a 1% CTB solution (List Biological Laboratories, Inc., Campbell, CA, USA) mixed with 2% dimethyl sulfoxide (DMSO) diluted in sterile water. …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt