Bacterial Toxin Research Citations

HEK293 cell transfection and stimulation:

… For TLR4/TLR5 co-transfections, HEK293 cells stably transfected with MD-2, CD14, and TLR4 were obtained from Invivogen (Catalog # 293-htlr4md2cd14). Cells were cultured in a 96-well flat-bottomed tissue culture plate at 50 10³ cells/well in cell growth media (DMEM, 4.5 g/l glucose, 50 U/ml penicillin, 50 g/ml streptomycin, 100 g/ml Normocin, 10 g/ml blasticidin, 50 g/ml Hygromycin B Gold, plus 10% fetal bovine serum (FBS) and 1% L-glutamine). The following day cells were transiently transfected using PolyFect transfection reagent (Qiagen, Valencia, CA) and the following DNA: NF-B-ELAM firefly luciferase 50 ng/well, HSV thymidine kinase-Renilla luciferase (pRL-TK, Promega) 5 ng/well, and huTLR5 (cloned into pEF6) and empty pEF6 vector at 110 ng/well as indicated. The transfection complexes were allowed to incubate with the cells for 24 hours. The following morning, the media in the wells was aspirated and the cells were then stimulated with ultrapure Escherichia coli O111:B4 LPS (List Biologicals, Campbell, CA) or ultrapure S. Typhimurium flagellin (Invivogen) suspended in cell growth media. After four hours, cells were lysed with passive lysis buffer and NF-B activation was determined as above.

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

… Microglial cells (3 10 4 ) were plated onto glass coverslips and treated with LPS (2.5 g/ml, List Biological Laboratories, #304) and IFN- (25 Units/ml, Sigma, #I4777) to stimulate and activate the cells (Klegeris and McGeer, 2001). …

• Product #304 – LPS from Salmonella minnesota R595 (Re)

… and PRN at 0.1 g (PT, cat. 180; FHA cat. 170; and PRN cat. 187; List Biological Laboratories)were determined by ELISA, as described by Quinello et al. [19], with some modifications. …

• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

Author did not specify which List Labs Pertussis Toxin was utilized.  List Labs provides Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer and Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free).

Product #170, Filamentous Hemagglutinin (FHA) from B. pertussis, has been discontinued.  Large quantities may be obtained through Bulk Requests and Customer Orders.

EAE induction, Treg administration, and CNS mononuclear cell isolation:

For EAE induction, 2-month-old Foxp3^Cre or Foxp1^f/fFoxp3^Cre mice were immunized subcutaneously with 200 µg of MOG3555 peptide (GL Biochem, Shanghai, China) emulsified in Complete Freunds Adjuvant (CFA; Sigma). Additionally, mice were injected intravenously with 200 ng of PT (List biological laboratories, Campbell, CA) on d0 and d2 following 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)

… Mice also received 100ng pertussis toxin (List Biological Laboratories) intraperitoneally 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)

EAE:

… The mice were intraperitoneally (ip) injected with 300 ng of pertussis toxin (List Biological Laboratories Inc., USA) at the same time of immunization and 48 h after 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)

… Cells cultured with only anti-CD28, anti-CD49d as negative controls (medium), and cultured with 1g/ml SEB (List Biological Labs, CA) served as positive controls. …

pTfh:B:APC cocultures:

Purified memory B, pTfh, and APCs (1 10⁴ cells each) were cocultured at a 1:1:1 ratio in duplicate wells in the presence of 5 g/mL of H1N1/09 vaccine Ag with 1 g/mL of anti-CD28 mAb for 7 d in the presence or absence of a cytokine cocktail containing IL-21 (50ng/ml), anti-IL-2 (10 g/ml) and antiTNF- (2 g/ml). Medium alone and B+APC served as negative controls, and 1 g/mL of SEB served as a positive control. …

… The brainstem was exposed via a limited occipital craniotomy and micropipettes containing the retrograde tracer cholera toxin subunit B conjugated to AlexaFluor 555 (CTB; 1% in deionized water, List Biological Laboratories, Campbell, CA) were inserted into the nTS at the level of calamus scriptorius: 0.4 mm lateral and 0.5 mm ventral from the surface. CTB was injected bilaterally (30 nl each side). …

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

EAE models:

Mice were immunized with MOG_35-55 (150 µg; Peptide International) emulsified in complete Freund’s adjuvant (CFA; Sigma-Aldrich) containing 10 mg/mL heatkilled Mycobacterium tuberculosis H37Ra (Difco Laboratories). The mice were injected intraperitoneally with pertussis toxin (List Biological Laboratories; 500 ng) on days 0 and 2. The adoptive transfer of encephalitogenic CD4⁺ T cells isolated 9 days after immunization was also used to induce EAE. …

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)

Lep-vesicle-recycling endosome-exocyst complex inhibition tests:

The cell monolayers were treated with 35 nM edema factor or lethal factor plus 70 nM protective antigen (EF + PA or LF + PA) of anthrax toxin (List Biological Laboratories, USA), the inhibitor (EF + PA) of Rab11 and inhibitor (LF + PA) of Sec15 to block Rab11-Sec15 binding, for 24 hr at 37C (Guichard et al., 2010), and then infected with L. interrogans strain Lai at MOI100 for 8 hr. The subsequent steps and detection of Lep-vesicle-Rab11-Sec15/Sec3 co-localization were the same as above. …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #178A – Anthrax Edema Factor (EF), Recombinant from B. anthracis
• Product #169 – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence