Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | One Shot Stbl3 Chemically Competent E .coli | Life Technologies | C7373-03 | |
| Strain, strain background (Escherichia coli) | DH10Bac bacteria | Thermo Fisher | 10361012 | |
| Cell line (Homo-sapiens) | 293FT, Human embryonic kidney | Invitrogen | R70007 | |
| Cell line (Homo-sapiens) | HEK293 GnTI--, Human embryonic kidney | ATCC | CRL-3022 | |
| Cell line (Homo-sapiens) | HEK293, Human embryonic kidney | ATCC | CRL-1573 | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor WT-MYC | Choi et al., 2016 | transfected construct (human) | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R14E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay, In vivo insulin-binding assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor P495A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor D496K-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor F497A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor D1120A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R498E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor K703A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor F714A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor K649E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor K652E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor Y477A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R479E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor K484E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R488E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor P536A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor P537A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor L552A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R554E-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor K484E,L552A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay, In vivo insulin-binding assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R345A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor E697A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay |
|
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor E695A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin receptor R14E,K484E,L552A-MYC | This paper | Materials and methods, subsection: Insulin receptor activation assay, In vivo insulin-binding assay | |
| Transfected construct (Homo-sapiens) | pCS2-human Insulin recetor Y960F, S962A, R1333A, I1334A, L1335A, L1337A-Myc | Choi et al., 2016 | transfected construct (human) | |
| Transfected construct (Homo-sapiens) | pEZT-BM-human Insulin receptor (Y960F, S962A, D1120N, R1333A, I1334A, L1335A, L1337A)−3C-Tsi3-His | This paper | Materials and methods, subsection: Insulin receptor activation assay, In vivo insulin-binding assay | |
| Transfected construct (Homo-sapiens) | pEZT-BM-human Insulin receptor (K484E, L552A, Y960F, S962A, D1120N, R1333A, I1334A, L1335A, L1337A)−3C-Tsi3-His | This paper | Materials and methods, subsection: Insulin receptor activation assay, In vivo insulin-binding assay | |
| Antibody | Rabbit monoclonal anti-IR-pY1150/1151 | Cell signaling | 19H7 | WB (1:1000) |
| Antibody | Mouse monoclonal anti-MYC | Roche | Clone 9E10 | WB (1:1000), IF (1:500) |
| Antibody | Anti-rabbit IgG (H+L) (Dylight 800 conjugates) | Cell signaling | 5151 | WB (1:5000) |
| Antibody | Anti-mouse IgG (H+L) (Dylight 680 conjugates) | Cell signaling | 5470 | WB (1:5000) |
| Commercial assay or kit | Micro BCA Protein Assay Kit | Thermo Scientific | 23235 | |
| Commercial assay or kit | Alexa Fluor 488 | Thermo Scientific | A10235 | |
| Commercial assay or kit | Q5 site directed mutagenesis kit | NEB | E0554S | |
| Commercial assay or kit | Gibson Assembly Master Mix | NEB | E2611 | |
| Chemical compound, drug | cOmplete Protease Inhibitor Cocktail | Roche | 5056489001 | |
| Chemical compound, drug | PhosSTOP | Roche | 4906837001 | |
| Chemical compound, drug | BMS536924 | Tocris | 4774 | |
| Software, algorithm | CytExpert 2.1 | Beckman Coulter | https://www.beckman.com/flow-cytometry/instruments/cytoflex/software | |
| Software, algorithm | Protparam | ExPASy | https://web.expasy.org/protparam/ | |
| Software, algorithm | Prism 7 | GraphPad | N/A | |
| Other | Sepharose 4B resin | GE Healthcare | 17012001 | |
| Other | Superose 6 Increase 10/300 GL | GE Healthcare | 29091596 | |
| Other | Superdex 75 increase 10/300 GL | GE Healthcare | 29148721 | |
| Other | Dynabeads Protein G | Invitrogen | 10003D | |
| Other | DMEM, high glucose | Thermo Fisher | 11965–118 | |
| Other | Pierce NHS-Activated Agarose, Dry | Thermo Fisher Scientific | 26197 | |
| Other | Amicon Ultra-15 Centrifugal Filter Units | Milliporesigma | UFC9100 | |
| Other | Amicon Ultra-0.5 Centrifugal Filter Units | Milliporesigma | UFC5100 | |
| Other | Quantifoil R 1.2/1.3 grid Au300 | quantifoil | Q37572 | |
| Other | ProLong Gold Antifade reagent with DAPI | Invitrogen | P36935 | |
| Other | Cellfectin | Invitrogen | 10362100 | |
| Other | Lipofectamine 2000 | Invitrogen | 11668019 | |
| Other | Human Insulin | Sigma | I2643 |