125i glp 1

Manufactured by PerkinElmer

Sourced in United States

125I-GLP-1 is a radioisotope-labeled form of the glucagon-like peptide-1 (GLP-1) molecule, which is used in research and development applications. It provides a tool for studying the interactions and properties of GLP-1 within experimental systems.

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Lab products found in correlation

Optiphase supermix, by PerkinElmer (2 mentions) Microbeta2 plate counter, by PerkinElmer (2 mentions) Filtermate harvester, by PerkinElmer (1 mentions) 125i glucagon, by PerkinElmer (1 mentions)

Spelling variants of this product

Radiolabeled 125I-GLP-1 (2 citations)

3 protocols using 125i glp 1

GIP and GLP-1 Receptor Binding Assay

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Example 7

Materials and Methods for GIP and GLP-1 Radioligand Competition Binding Assay

Membranes were prepared from mammalian cell lines overexpressing hu GIPR and hu GLP-1R and were typically stored at −80° C. in Assay Buffer: 50 mM HEPES (7.4), 5 mM MgCl₂, 1 mM CaCl₂, 0.2% BSA and Protease Inhibitor Tablets “EDTA Free”. Unlabeled GIP and GLP-1 peptides were obtained from Phoenix Pharmaceuticals and Radiolabeled ¹²⁵I-GIP and ¹²⁵I-GLP-1 were ordered from Perkin Elmer. Primary reactions were conducted for 2 h at room temperature with shaking in 96 well cell culture round bottom plates (Costar). Cell membrane concentration was fixed at 0.6 mg/mL and radioligands were used at a concentration of 0.1 nM. Test articles were titrated at six concentrations in duplicate. The primary plate was harvested after two hours with ice cold 50 mM TRIS (pH 7.4) using a Filtermate Harvester (Perkin Elmer) and cell membranes were captured on PEI-treated GF/C UniFilter plates (Perkin Elmer). Plates were dried for 15 min using a vacuum oven and Microscint 20 was added to each well before counting in a Perkin Elmer TopCount NXT HTS instrument. Specific binding was determined by taking the difference of total and non-specific counts. Non-specific wells contained 1 μM cold ligand. Data was analyzed and plotted using GraphPad Prism 7 as shown in FIG. 22.

US10905772B2. Method of treating or ameliorating metabolic disorders using GLP-1 receptor agonists conjugated to antagonists for gastric inhibitory peptide receptor (GIPR) (2021-02-02). AMGEN INC. [US]. Inventors: Yuan Cheng [US], Chawita Netirojjanakul [US], Jerry Ryan Holder [US], Bin Wu [US], James R. Falsey [US], Bradley J. Herberich [US], Kelvin Sham [US], Leslie P. Miranda [US], Shu-Chen Lu [US], Murielle M. Veniant-Ellison [US], Shanaka Stanislaus [US], Junming Yie [US], Jing Xu [US].

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Homogeneous Binding Assay for GLP-1R

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CHO-K1 cells stably expressing WT or mutant GLP-1R were seeded into 96-well plates at a density of 3 × 10⁴ cells/well and incubated overnight at 37 °C and 5% CO₂. For homogeneous binding, cells were washed twice and incubated with a blocking buffer (F12 supplemented with 33 mM HEPES, and 0.1% (w/v) BSA, pH 7.4) for 2 h at 37 °C. Then, radiolabeled ¹²⁵I-GLP-1 (40 pM, PerkinElmer, Waltham, MA, USA) and unlabeled compounds were added and reacted with the cells in binding buffer (F12 supplemented with 33 mM HEPES, and 0.1% (w/v) BSA, pH 7.4) at 4 °C overnight. Cells were washed three times with ice-cold PBS and lysed using 50 μL of lysis buffer (PBS supplemented with 20 mM Tris–HCl and 1% Triton X-100, pH 7.4). Subsequently, the plates were counted for radioactivity (counts per minute, CPM) in a scintillation counter (MicroBeta² Plate Counter, PerkinElmer, Waltham, MA, USA) using a scintillation cocktail (OptiPhaseSuperMix; PerkinElmer, Waltham, MA, USA).

Zhou Q., Guo W., Dai A., Cai X., Vass M., de Graaf C., Shui W., Zhao S., Yang D, & Wang M.W. (2021). Discovery of Novel Allosteric Modulators Targeting an Extra-Helical Binding Site of GLP-1R Using Structure- and Ligand-Based Virtual Screening. Biomolecules, 11(7), 929.

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Radioligand Binding Assay for GLP-1R and GCGR

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CHO cells stably expressing GLP-1R or GCGR were seeded into 96-well plates at a density of 3 × 10⁴ cells/well and incubated overnight at 37 °C and 5% CO₂. The radioligand binding assay was performed 24 h thereafter. For homogeneous binding, cells were incubated in binding buffer with a constant concentration of ¹²⁵I-GLP-1 (40 pM, PerkinElmer, Boston, MA) or ¹²⁵I-glucagon (40 pM, PerkinElmer) and unlabeled compounds at 4 °C overnight. Cells were washed three times with ice-cold PBS and lysed using 50  μl lysis buffer (PBS supplemented with 20

m M

Tris–HCl and 1% Triton X-100, pH 7.4). Subsequently, the plates were counted for radioactivity (counts per minute, CPM) in a scintillation counter (MicroBeta2 Plate Counter, PerkinElmer) using a scintillation cocktail (OptiPhase SuperMix; PerkinElmer).

Wan F., Zhu Y., Hu H., Dai A., Cai X., Chen L., Gong H., Xia T., Yang D., Wang M.W, & Zeng J. (2020). DeepCPI: A Deep Learning-based Framework for Large-scale in silico Drug Screening. Genomics, Proteomics & Bioinformatics, 17(5), 478-495.

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