Bacterial Toxin Research Citations

Tracer injections:

Males and females MC4-R-GFP mice were deeply anesthetized with ketamine (5 mg/100 g) and xylazine (1 mg/100 g). The animals (n=10 males, n=4 females) received unilateral stereotaxic injections of the retrograde tracer cholera toxin b subunit (CTb 1%; List Biological Laboratories) into the PVH (0.43 mm from bregma; +0.23 lateral; 4.75 mm from surface of the skull). The retrograde tracer was injected with a glass micropipette and air pressure injection system as described previously …

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

… Injection site, gt. CTb, 1:50,000, List Biological Laboratories, Campell, CA, …

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

Experimental autoimmune encephalomyelitis (EAE)

Age- (5-8 weeks) and sex-matched mice were immunized subcutaneously with 100 g MOG₃₅₅₅ peptide (MEVGWYRSPFSRVVHLYRNGK) emulsified in CFA (Difco Laboratories) with 500 g Mycobacterium tuberculosis on day 0. Mice also received 200 ng pertussis toxin (List Biological Laboratories) by intraperitoneal injection 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)

ET Assays.

EF was obtained from List Biological Laboratories. J774A.1 cells in 384-well plates were incubated with 11 nM PA and 3.7 nM EF for 5 h at 37 C, and the concentration of cAMP was measured by cAMP enzyme immunoassay…

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).

Materials and Methods:

…Coronal sections of 40 m were cut on a freezing microtome, after which the sections were post-fixed in 4% PFA at 4C for 4 hours. Free-floating sections were then washed several times in 0.5% PBST (PBS tween), and then washed several times in 2.5% normal rabbit serum (diluted in 0.5% PBST). Sections were incubated in goat anti-CTB (#703, List Biological Laboratories, Campbell, CA) at 14000 in 2.5% normal rabbit serum diluted in 2% PBST for 96 hours at 4C. …

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

Ag and adjuvant:

… CT was purchased from List Biological Laboratories (Campbell, CA, USA).

Immunization and sample collection

Mice were immunized nasally with a 20-μL aliquot (10 μL per nostril) of phosphate-buffered saline (PBS) containing 10 μg 40k-OMP alone or combined with 1 μg CT on days 0, 7, and 14. Serum, saliva, and nasal wash samples were collected, as described elsewhere (21), to examine 40k-OMP-specific Ab responses.

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

ELISA and western blot:

The cross-reactivity of scFv 2LF with EF was tested by ELISA. Briefly, microtiter plates were coated with 2 g/ml of EF (List Biological Laboratories, Campbell, CA), while LF (List biological Laboratories) and KLH (Sigma-Aldrich, Saint-Quentin Fallavier, France) were utilized as positive and negative controls respectively. …

Affinity measurements:

Affinity constants (K_D) and dissociation constants (k_d) of scFv 2LF for LF, EF and their variants were measured by surface plasmon resonance (SPR) with a Biacore X (Biacore, Uppsala, Sweden) instrument. …

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).

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).

Anterograde and retrograde tracing:

Three weeks later, we perfused the mice and cut brain sections as above. For immunostaining, we placed sections in 0.3% hydrogen peroxide for 30 min and then incubated them overnight in primary antibody at room temperature. Primary antibodies included goat anti-CTB (1:15,000; List Biological),

For the combination of VGAT in situ hybridization and CTB immunostaining, we incubated sections with goat anti-CTB antibody overnight, followed by Alexa Fluor 488-conjugated anti-goat IgG antibody (1:1000; Invitrogen) for 1 h.

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

EAE induction and treatment:

Mice were immunized subcutaneously at the base of the neck and tail with 50 g of PLP₁₃₉₁₅₁ (C140S) peptide in incomplete Freund’s adjuvant (IFA) (Sigma) containing 400 g of Mycobacterium tuberculosis H37Ra (Difco Laboratories- Detroit USA). One day after priming, mice were injected intravenously with 200 ng of Pertussis toxin (List Biological Laboratories, Inc UK) and scored daily for clinical signs of EAE, according to a standard scale to monitor the severity of EAE course [26]. …

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)

Reagents:

…SNAPtide (FITC/DABCYl, product no. 521) fluorogenic peptide was from List Biological Laboratories (Campbell, CA)…

Substrates and BoNT/A subtypes:

To determine if the BoNT/A-cleavable peptides were hydrolyzed by BoNT/A subtypes, we exposed combinations of cleavable substrates and non-cleavable controls (fN with mA, and mN with fA), and commercial SNAPtide to the LCs of subtypes A1A5 (Figure 1C). fN, mN, and SNAPtide were cleaved by LC-A1, A2, A3, and A5, but not by LC-A4. LC-A4, however, did cleave recombinant SNAP-25, albeit to a much lesser extent than did A1-3 and A5, indicating that LC-A4 had catalytic activity…

• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin