Citations

Ablation of AgRP neurons

To ablate AgRP neurons, mice carrying the AgrpDTR/+ allele, were bilaterally microinjected with DT (0.4 ng/side/mouse, List Biological Laboratories, Campbell, CA) using a needle (Hamilton Small Hub RN 33 G, Reno, NV) connected with a 10 μl syringe (Hamilton 700 Microliter, Reno, NV), at a rate of 0.1 μl/min. A total 0.4 μl volume/side was delivered into the dlDRN2. To describe the expression profile of AgRPARC-dlDRN neurons, the vehicle (0.9% NaCl) or DT were injected into the dlDRN with AgrpDTR/+::NpyGFP mice. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

In vivo retinal monitoring in EAE

Mice. For experiments involving in vivo retinal imaging, CX3CR1GFP/+–knock-in mice were fully backcrossed into C57BL/6J. We confirmed the absence of an RD8 mutation (associated with hereditary macular degeneration). In CX3CR1GFP/+–knock-in mice, microglia, macrophages, monocytes, and NK cells express GFP, which can be imaged through CSLO (14), without any immunologic alterations (15).

Induction and clinical scoring of MOG35-55 EAE. Seven female mice at 10 weeks of age were immunized s.c. with 100 μg of MOG35–55 peptide (Genemed Synthesis) in complete Freund’s adjuvant containing 400 μg Mycobacterium tuberculosis H37Ra (Difco Laboratories). Mice received 200 ng pertussis toxin (List Biological) by i.p. injection at the time of and after immunization. We monitored clinical scores, as described before (42). …

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)

… anthrax protective antigen (recombinant from Bacillus anthracis; 0.5 µg/ml; List Biological Laboratories, Inc.), …

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

Induction and clinical evaluation of EAE

We induced EAE by immunization of mice with 4 mg/ml MOG35–55 peptide (TS-M704-P; MBL, Nagoya, Japan) emulsified in complete Freund’s adjuvant containing 1 mg/ml Mycobacterium tuberculosis H37RA (#231131; BD Difco, Lawrence, KS, USA) at a dose of 200 μg per mouse, followed by intraperitoneal injections of 300 ng pertussis toxin (# 180-A1; List Biological Laboratories Inc., Campbell, CA, USA) per mouse on days 0 and 2. …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

2.2. Induction and evaluation of EAE

The EAE inducing antigen employed was myelin oligodendrocyte glycoprotein peptide (MOG35-55). To induce EAE, complete Freund’s adjuvant (CFA) emulsified with an equal volume of phosphate-buffered saline (PBS) containing 250 µg of Mycobacterium tuberculosis strain H37Ra (BD Biosciences, San Jose, CA, USA) and 200 µg of MOG35–55 (MEVGWYRSPFSRVVHLYRNGK) (Wuhan Bioyears gene Biotechnology Inc., China), was injected into the mice subcutaneously in each axillary fossa, on experimental day 0 and day 2, immediately followed by an intravenous injection of pertussis toxin (200 ng, List Biological Laboratories, Inc., Campbell, CA, USA) in the tail vein. The control group received only CFA in place of the EAE induction solution. …

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 received 200 ng of pertussis toxin (List Biological Laboratories) IP on day 0 (immunization day) and day 1 (24 hours 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)

2.2 Drugs

… Diphtheria toxin (List Biological Laboratories) was dissolved in saline and administered at 50 mg/kg, based on previous reports …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

2.3. LT Neutralization Activity (LTNA)

Recombinant PA and LF were purchased from List Biological (List Biological Laboratories Inc., Campbell, CA, USA). The ability of participant plasma to neutralize LT (LTNA) was adapted from the standard procedure from the CDC as previously described [29] using the J774.1 macrophage cell line (ATCC, Manassas, VA, USA). In brief, cells were plated overnight at 90,000 cells/well in a 96-well plate. Plasma samples were diluted 1:50 in culture medium, and then diluted for eight additional serial two-fold dilutions in dilution tubes, and then incubated with LT for 30 min at 37 °C (600 ng/mL of PA, 120 ng/mL of LF) before adding to cell cultures. Control wells included cells alone, cells with added PA only, LF only, or cells with PA and LF (LT). …

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

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

EAE Induction

Mice were injected s.c. at the tail base with an emulsion containing equal amounts of complete Freund’s adjuvant (CFA) (Difco) containing 5 mg/ml Mycobacterium tuberculosis (strain H37Ra) (Difco) and 200 µg MOG35-55 in PBS. 400 ng of pertussis toxin (List Biological Labs) was injected i.p. on days 0 and 2 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)

2.1. Materials and Reagents

… Diphtheria toxoid (DT), pertussis toxin (PT), and TT used as coating antigens for ELISA were purchased from List Biological Laboratories, Inc. (Campbell, CA, USA). …

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae
• Product #190 – Tetanus Toxin from Clostridium tetani

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)