Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | BL21(DE3) | Sigma-Aldrich | CMC0016 | Electrocompetent cells |
| Strain, strain background (Escherichia coli) | MC1061 | J Biol Chem. 261:13844–9. PMID:3531212 | NA | Electrocompetent cells |
| Other | Protein-G-agarose | Roche | 11243233001 | Resin used for immunoprecipitation |
| Antibody | Anti-HA.11 epitope tag antibody (mouse monoclonal) IgG | BioLegend | Cat# 901533 | Used for immunoprecipitation (1 μl of 1 mg/ml, diluted 1:820) |
| Antibody | LepB antibody (rabbit polyclonal) IgG |
Generated in-house | NA | Used for immunoprecipitation (dilution 1:820) |
| Recombinant DNA reagent | pET Duet-1 (plasmid) | Novagen | Cat# 71146 | Expression plasmid |
| Recombinant DNA reagent | pING1 (plasmid) | Gene 34:137–45. PMID:4007491 | NA | Expression plasmid |
| Commercial assay, kit | GeneJET Plasmid miniprep kit | Thermo Fisher Scientific RRID: SCR_008452 |
Cat# 0502 | Used to purify plasmids |
| Commercial assay, kit | GeneJET PCR Purification Kit | Thermo Fisher Scientific |
Cat# K0701 | Used to purify linear fragments for in vitro expression |
| Commercial assay, kit | PURExpress | New England Biolabs | Cat# E6800L | Used for in vitro expression |
| Chemical compound, drug | 35S methionine | PerkinElmer | Cat# NEG009T001MC |
35S Methionine is incorporated into the protein during in vitro and in vivo translation and aids detection by phosphor imaging |
| Software, algorithm | EasyQuant | Developed in-house Nat Struct Mol Biol. 19:1018–22. PMID: 23001004 |
Used to quantify relative fraction full length of translated protein from SDS-PAGE |