| Targets |
Semaglutide, FITC labeled targets the glucagon-like peptide-1 receptor (GLP-1R). Semaglutide is a potent and selective GLP-1R agonist. Upon binding to GLP-1R, which is a G-protein-coupled receptor (GPCR), it activates the cAMP signaling pathway, leading to increased insulin secretion, decreased glucagon secretion, slowed gastric emptying, and appetite suppression. The FITC label does not interfere with receptor binding, allowing the fluorescent conjugate to be used as a tracer for receptor localization and internalization studies.
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| ln Vitro |
In vitro, Semaglutide, FITC labeled is used to study GLP-1 receptor binding and internalization. In a typical binding assay, cells expressing GLP-1R (e.g., INS-1E beta-cells or HEK293-GLP-1R) are incubated with the labeled peptide (10-1000 nM) for 1-2 hours at 4degC (to prevent internalization). After washing, bound fluorescence is measured by flow cytometry or fluorescence microscopy. The signal is specific, as it can be competed away by excess unlabeled semaglutide. This allows for quantitative determination of binding affinity (Kd).
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| ln Vivo |
In vivo, Semaglutide, FITC labeled is a research tool for visualizing GLP-1R distribution. It can be injected intravenously into mice to assess receptor binding in various organs (e.g., pancreas, brain). For example, after intravenous injection (0.1-1 mg/kg), mice are euthanized after 1-4 hours, and organs are harvested, sectioned, and imaged by fluorescence microscopy. Specific binding can be blocked by pre-injection with excess unlabeled semaglutide. This technique is used to study GLP-1R expression patterns in normal and disease states.
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| Enzyme Assay |
Non-cellular assays are not typically performed. However, the dye is characterized by UV-Vis spectroscopy. A solution of Semaglutide, FITC labeled (10 ug/mL in PBS) is scanned from 300-600 nm. The FITC label shows a characteristic absorption peak at ~495 nm, and the peptide shows an absorption peak at ~280 nm. The degree of labeling (DOL) is calculated from the ratio of absorbance at 495 nm to that at 280 nm. A DOL of 0.5-1.0 is typical for a functional probe.
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| Cell Assay |
For a cell binding assay, GLP-1R-expressing cells (e.g., INS-1E) are seeded into 96-well plates at 2×10⁴ cells/well. After 24 hours, cells are incubated with Semaglutide, FITC labeled (100 nM) in binding buffer (PBS + 0.1% BSA) for 1 hour at 4degC. Cells are washed three times with cold PBS, and fluorescence is measured using a microplate reader (Ex 485 nm, Em 535 nm). To confirm specificity, a parallel set of wells is pre-incubated with 10 uM unlabeled semaglutide (blocking). The signal is reduced by >80% in the blocked wells, confirming specific binding to GLP-1R.
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| Animal Protocol |
For receptor internalization studies, GLP-1R-expressing cells are incubated with Semaglutide, FITC labeled (100 nM) at 37degC for 0, 5, 15, 30, 60 minutes. The cells are then fixed and counterstained with DAPI to label nuclei. Confocal microscopy images show the labeled peptide initially on the cell surface and progressively accumulating in cytoplasmic vesicles. This demonstrates receptor-mediated endocytosis. In vivo imaging studies in mice are performed using an IVIS spectrum imager after intravenous injection of the fluorescent probe (Ex 495 nm, Em 520 nm).
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| ADME/Pharmacokinetics |
The parent peptide, semaglutide, has a molecular weight of 4113 g/mol, while the FITC-labeled version has a molecular weight of ~4500 g/mol. Semaglutide is a long-acting GLP-1 analog with a plasma half-life of approximately 1 week in humans due to its albumin-binding fatty acid side chain. The FITC-labeled version is expected to have a similar PK profile if administered in vivo. It is stable in plasma for several hours. Storage: lyophilized powder at -20degC, protected from light. In solution, store at -80degC in small aliquots.
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| Toxicity/Toxicokinetics |
Semaglutide, FITC labeled is a research chemical and is not for human use. The parent drug semaglutide is an FDA-approved drug (Ozempic®, Wegovy®) with known side effects including gastrointestinal disturbances (nausea, vomiting, diarrhea) and, rarely, pancreatitis or medullary thyroid carcinoma risk. The labeled version should be handled with the same precautions as a potent bioactive peptide. Standard lab safety: use gloves, lab coat, and goggles.
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| References | |
| Additional Infomation |
Semaglutide, FITC labeled (CAS 2241669-86-1) is a valuable tool for studying the trafficking and localization of the GLP-1 receptor. Semaglutide is a long-acting GLP-1 analog used clinically for type 2 diabetes (Ozempic) and obesity (Wegovy). The fluorescent conjugate allows visualization of receptor binding in real-time. This compound is for research use only and is not for therapeutic or diagnostic use. Purity: peptide content ≥60%, FITC content determined by HPLC.
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| Appearance |
Typically exists as solids at room temperature
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~6.67 mg/mL (~1.48 mM; with ultrasonication)
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| Solubility (In Vivo) |
Note: Please refer to the "Guidelines for Dissolving Peptides" section in the 4th page of the "Instructions for use" file (upper-right section of this webpage) for how to dissolve peptides. Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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