Citations

HPLC Method to Assay the ADP – Ribosyltransferase Activity of Cholera Toxin

Materials: Cholera Toxin (Prod # 100B) and CTB (Prod # 104) are products of List Biological Laboratories, Inc.

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae
• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

Materials:

The endotoxin (LPS) from E. coli K12 LCD25 was purchased from List Biological Laboratories. All LPS preparations from List Biological Laboratories, Inc., are essentially free of nucleic acid and protein and are chemically characterized with respect to their phosphate and 2-keto-3-deoxyoctonate (KDO) contents. For these studies, the purchased LPS preparation was further purified by a phenol re-extraction method to eliminate residual protein contamination prior to use in our experimental models. …

• Product #314 – LPS from Escherichia coli K12 strain LCD25

In vitro characterization of plant-produced mAbs:

ELISA was performed as previously described in reference 36, with slight modifications. Plates were coated with 1 g/mL of recombinant B. anthracis PA (List Biological Laboratories Inc., Campbell, CA). …

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

Materials:

SNAPtide substrates (Products #520 and #521), the Control Peptide for SNAPtide #520 (Product #525) and Botulinum neurotoxin type A light chain (Product #610A) are products of List Biological Laboratories, Inc.

• Product #520 – SNAPtide Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin
• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin
• Product #525 – Control Peptide for SNAPtide 520 Substrate for C. botulinum Type A Neurotoxin
• Product #610A – Botulinum Neurotoxin Type A Light Chain, Recombinant

Methods – The above materials were used in the following (refer to poster for details):

• Fluorimetric assays
• Determination of IC₅₀ and mode of inhibition

Materials:

SNAPtide  substrate (Product #520) and Botulinum neurotoxin type A (Product #130A) are products of List Biological Laboratories, Inc.

• Product #520 – SNAPtide Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin
• Product #130A – Botulinum Neurotoxin Type A from Clostridium botulinum

Methods – The above materials were used in the following (refer to poster for details):

• BoNT/A capture using the F1-2 monoclonal antibody
• BoNT/A capture using the F1-40 monoclonal antibody

CYTOKINE PRODUCTION ASSAY AND MEASUREMENT OF INTERLEUKIN-1 AND TNF-:

… LPS (List Biological Laboratories, cat #201) was diluted in assay medium. Assay medium consisted of RPMI 1640 (Invitrogen Life Technologies Corporation, cat #11875) containing 1 penicillin/streptomycin/glutamine (Invitrogen, cat #10378) and 0.1% human serum albumin (Bayer AG, cat #69220). Cultures of 50% whole blood/50% assay medium in a final volume of 200 L were incubated for 18 h at 37 C and 5% CO₂ in the presence of 0 or 100 g/L LPS in a 96-well tissue-culture plate (BD Biosciences, cat #353072). LPS stimulation was initiated <1 h after blood draw and samples were run as triplicates. …

• Product #201 – LPS from Escherichia coli O111:B4

Cell culture and infection experiments:

RAW264.7 macrophage-like cells (ATCC No. TIB-71) were cultured in RPMI (10% FCS, 1% L-Glutamine) at 37C/5% CO₂. For infection experiments for analysis of LDH and Il-1 release, cells were seeded in 96 well plates the day before. 20000 RAW264.7 cells were seeded per well and were pre-stimulated with 1 g/ml E. coli LPS (List Biological Laboratories, California) overnight to induce expression of pro-IL-1. …

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

 Materials and Methods:

… The light-chain activity and cleavage pattern of these BoNT/A products were assessed using the novel LCA-HPLC method reported here with BoNT/A substrate (SNAPtide520; List Biological Laboratories, Inc.; Campbell, CA, USA). SNAPtide520 sequence: Abz Thr D-Arg Ile Asp Gln Ala Asn Gln Arg Ala Thr Lys Nle Lys Dnp; where Abz = o-Aminobenzoic acid (FRET pair fluorophore) and Dnp = 2,4-Dinitrophenyl (FRET pair quencher).

BoNT/A cleavage fragment (designated as SNAPtide529 by List Biological Laboratories, Inc.) was utilized to identify retention time of the fragment. …

Commercially available BoNT/C FRET substrate (SYNTAXtide560; List Biological Laboratories, Inc.; Campbell, CA, USA) was utilized to assess type-C cleavage. Because of the RP-HPLC reaction-product readout, no modification to the method was necessary in order to observe the reaction products generated using the SYNTAXtide560 substrate. …

• Product #520 – SNAPtide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin
• Product #529 – Unquenched Calibration Peptide for SNAPtide 520 Substrate for C. botulinum Type A Neurotoxin
• Product #560 – SYNTAXtide (o-Abz/Dnp) Peptide Substrate for C. botulinum Type C Neurotoxin

Protein Purification:

Highly purified recombinant proteins were prepared in our laboratories for binding assays as well as immunizations, as described below. For ELISA and LTx neutralization assays, purified PA and LF were purchased from List Biologicals Laboratories, Inc., 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).

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

PTx (holotoxin), the full length PTx-S1 (1-235) protomer, and the B oligomer were purchased from List Biological Laboratories, Inc., Campbell, CA.

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)