Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | CaSR | NCBI | NM_000388.4 | |
| Strain, strain background (Escherichia coli) | BL21(DE3) | New England Biolabs | Cat#: C2527I | E. coli strain for expression of the nanobody |
| Strain, strain background (Escherichia coli) | TG1 | Lucigen | Cat#: 60,502 | Electrocompetent cells |
| Strain, strain background (Escherichia coli) | TOP10F' | Huayueyang Biotech | WXR15-100S | E. coli strain for expression of the nanobody |
| Cell line (Homo sapiens) | (HEK) 293 S GnTI- cells | ATCC | Cat# CRL-3022RRID: CVCL_A785 | Mycoplasma negative |
| Cell line (Homo sapiens) | HEK 293T/17 | ATCC | Cat# CRL-11268RRID: CVCL_1926 | Mycoplasma negative |
| Antibody | HA-Tag, (Mouse monoclonal) | Yeasen | 30,701ES60 | Dilution: (1/2000) |
| Antibody | Flag-Tag (DYKDDDDK), (Mouse monoclonal) | Yeasen | 30,503ES20 | Dilution: (1/2000) |
| antibody | Peroxidase AffiniPure Goat Anti-Mouse IgG (H + L)(Goat monoclonal) | Yeasen | 33,201ES60 | Dilution: (1/2500) |
| Recombinant DNA reagent | AxyPrep Plasmid Miniprep Kit | CORNING LIFE SCIENCES | Cat#:220 | |
| Recombinant DNA reagent | pMECS vector | BioVector NTCC | pMECS | Phage display vector |
| Recombinant DNA reagent | pEG BacMam vector | Addgene | Cat#:160,451 | Vector |
| Recombinant DNA reagent | pCMV-HA | Addgene | Cat#:631,604 | Vector |
| Recombinant DNA reagent | pcDNA3.1 | Addgene | Cat#:128,034 | Vector |
| Peptide, recombinant protein | Flag peptide | Genscript | DYKDDDDK | |
| Peptide, recombinant protein | NB88(camel nanobody) | This study | Isolated from phage display library of immunized cammel with hCaSR | |
| Peptide, recombinant protein | NB-2D11 (camel nanobody) | This study | Isolated from phage display library of immunized cammel with hCaSR | |
| Commercial assay or kit | Luciferase assay kit | Promega | E152A | For signaling assay |
| Commercial assay or kit | SuperSignal ELISA Femto Substrate | Thermo Scientific | Cat#: 37,075 | Protein Assays and Analysis |
| Commercial assay or kit | Fluo-4, AM, Cell Permeant | YEASEN | 40,704ES50 | |
| Chemical compound, drug | TNCA | aladdin | 42438-90-4 | |
| Chemical compound, drug | NPS-2143 | aladdin | 284035-33-2 | |
| Chemical compound, drug | cinacalcet | aladdin | 364782-34-3 | |
| Chemical compound, drug | Lauryl Maltose Neopentyl Glycol (LMNG) | Anatrace | NG310 | Membrane protein purification |
| Chemical compound, drug | Glyco-Diosgenin (GDN) | Anatrace | GDN101 | Membrane protein purification |
| Chemical compound, drug | Cholesterol Hemisuccinate tris Salt (CHS) | Anatrace | CH210 | Membrane protein purification |
| Chemical compound, drug | TMB substrate | Thermo Fisher Scientific | 34,021 | Protein Assays and Analysis |
| Software, algorithm | cryoSPARC | https://cryosparc.com | Version 3.0.0RRID:SCR_016501 | Cryo-EM data processing |
| Software, algorithm | PHENIX | http://www.phenix-online.org/ | Version 1.19.2RRID:SCR_014224 | Structure refinement |
| Software, algorithm | Coot | Coot (cam.ac.uk) | Version 0.9.4 RRID:SCR_014222 | Structure refinement |
| Software, algorithm | MolProbity | DOI:10.1107/S0907444909042073 | RRID:SCR_014226 | Structure verification |
| Software, algorithm | UCSF Chimera |
https://wwwcgl.ucsf.edu/chimera/ (PMID:15264254) |
Version 1.15 RRID:SCR_004097 | Initial homology model docking |
| Software, algorithm | PyMol | Schrodinger | Version 2.5 RRID:SCR_000305 | Structural visualization/figure preparation |
| Software, algorithm | GraphPad Prism 7 | GraphPad | RRID:SCR_002798 | Analysis of signaling data |
| Other | Lipofectamine 2000 | Invitrogen | 11668030 | Transfection reagent for signaling assay |