Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| gene (Leptospira interrogans
serovar Pomona) |
LigB (1-12) | GenBank | GenBank:FJ030916 | |
| recombinant DNA reagent | pET28-His-Sumo (plasmid) | other | Re-engineered by Dr. Mao's lab (Manford et al., 2010) |
|
| strain, strain background (Escherichia coli) |
E. coli Rosetta strain | Novagen | Novagen:70954 | |
| commercial assay or kit | Ni-NTA resin | Qiagen | ||
| peptide, recombinant protein | Sumo protease (Ulp-1) | other | protein expression by Dr. Chang's lab utilizing pET28-His- Sumo vector provided by Dr. Mao's lab, and IPTG induction in Rosetta strain E. coli |
|
| commercial assay or kit | Superdex75 (size exclusion chromatography) |
GE Healthcare | GE_Life_Sciences: 17517401 |
|
| software, algorithm | BeStSel | DOI:10.1073/pnas.1500851112 | Uses CD spectra obtained from Aviv-201 spectropolarimeter |
|
| software, algorithm | Clustal Omega | DOI:10.1093/nar/gkt376 | RRID:SCR_001591 | |
| software, algorithm | ATSAS Suite (GNOM; DAMMAVER; DAMMIF; AMBIMETER) |
DOI:10.1107/S1600576717007786 | RRID:SCR_015648 | |
| software, algorithm | RAW | DOI:10.1107/S1600576717011438 | ||
| biological sample (L. interrogans) |
LigB1-7 | this lab | protein expression utilizing pET28-His- Sumo vector and IPTG induction in Rosetta strain E. coli |
|
| biological sample (L. interrogans) |
LigB7-12 | this lab | protein expression utilizing pET28-His- Sumo vector and IPTG induction in Rosetta strain E. coli |
|
| biological sample (L. interrogans) |
LigB5-12 | this lab | protein expression utilizing pET28-His- Sumo vector and IPTG induction in Rosetta strain E. coli |
|
| commercial assay or kit | Protein A/G Chromatography |
ThermoFisher Scientific Pierce | ||
| cell line (Mus musculus) | hybridoma clones | William Davis Laboratory | mAb-producing cell lines generated from BALB/c mice immunized with recombinant LigB antigen |
|
| antibody | HRP-conjugated anti-mouse IgG (goat polyclonal) |
Invitrogen | RRID:AB_2533947 | (1:5000) |
| antibody | anti-LigB1-7 (mouse monoclonal) |
this lab | (1:500) | |
| antibody | anti-LigB7-12 (mouse monoclonal) |
this lab | (1:500) | |
| antibody | HRP-conjugated anti-hamster IgG goat polyclonal) |
KPL | https://www.seracare.com/products/kpl-antibodies-and-conjugates/secondary-antibodies/anti-hamster-igg--h-l--antibody/ | (1:1000) |
| antibody | HRP-conjugated anti-hamster IgM goat polyclonal) |
SouthernBiotech | https://www.southernbiotech.com/?catno=6060-05&type=Polyclonal | (1:1000) |
| antibody | FITC-conjugated anti-mouse IgG (goat polyclonal) |
ThermoFisher Scientific | RRID:AB_2533946 | (1:1000) |
| antibody | isotypic control IgG (mouse Fc fragment) |
ThermoFisher Scientific | RRID:AB_10959891 | |
| strain, strain background (Leptospira biflex) |
Leptospira biflex | other | Colony maintained in Dr. Chang's lab |
|
| strain, strain background (L. interrogans) |
Leptospira interrogans
serovar Pomona |
this lab | Colony maintained in Dr. Chang's lab |
|
| strain, strain background (L. interrogans) |
Leptospira interrogans
serovar Manilae M1307 |
other | Colony maintained in Dr. Gerald Murray and Dr. Ben Adler's labs, provided as a gift |
|
| biological sample (Homo sapiens sapiens) |
Normal human serum | ImmunoReagents | ImmunoReagents: SP-001-VX10 |
|
| biological sample (Mesocricetus auratus) |
Golden Syrian Hamster | Harlan Sprague Dawley Laboratory |
5 weeks old at initial subcutaneous vaccination |
|
| chemical compound, drug | Adjuvant 2% Alhydrogel | InvivoGen |