As a fluorescent probe rather than a therapeutic agent, FITC-Ureidopropionic acid does not exert biological activity by binding to specific pharmacological targets. The probe is designed to utilize ureidopropionic acid as a carrier molecule. Ureidopropionic acid is an endogenous metabolite involved in pyrimidine and amino acid metabolism, serving as an intermediate in the uracil degradation pathway where it is converted from dihydrouracil by dihydropyrimidinase and subsequently to β-alanine by β-ureidopropionase . Thus, the probe can theoretically be used to study the biodistribution and metabolic fate of this metabolite.
FITC-Ureidopropionic acid does not bind to a specific biological receptor. It functions as a fluorescent probe for labeling biomolecules or cells. Ureidopropionic acid is a naturally occurring metabolite in the body, involved in amino acid and pyrimidine metabolism. The FITC label provides bright green fluorescence upon excitation, enabling detection. The probe can be used to track ureidopropionic acid-associated processes or to label proteins via the carboxylic acid group.

FITC-Ureidopropionic acid is not designed to exert direct pharmacological activity (e.g., enzyme inhibition or receptor activation). Its primary in vitro application is as a fluorescent labeling reagent for biomolecules and cells. The FITC moiety provides bright green fluorescence with good photostability when excited at 490 nm and detected at 520 nm . The probe's activity is defined by its fluorescence signal intensity, which can be quantified via fluorescence spectrophotometry, flow cytometry, or fluorescence microscopy . The small molecular size of the probe facilitates cellular uptake, enabling intracellular tracking applications .

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FITC-Ureidopropionic acid is not studied for independent bioactivity such as enzyme inhibition or receptor binding. As a fluorescent probe, its primary utility is in imaging and flow cytometry applications. It can be used to label biomolecules or cells, allowing researchers to visualize the uptake, localization, and metabolism of ureidopropionic acid or to track ureidopropionic acid-conjugated molecules. The fluorescent signal is bright and photostable.

FITC-Ureidopropionic acid lacks traditional in vivo pharmacodynamic activity as it is not a therapeutic compound. However, it serves as a valuable tool for in vivo tracking studies. The fluorescent label allows visualization of the probe's distribution, accumulation, and potential metabolic transformation in living organisms. The ureidopropionic acid component can theoretically direct the probe to tissues or cells involved in pyrimidine metabolism . No specific therapeutic efficacy data is available, as the probe is exclusively for research applications.

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In vivo imaging studies with FITC-Ureidopropionic acid are limited. As a fluorescent probe, it may be used for ex vivo tissue imaging or for in vivo imaging in transparent models (e.g., zebrafish embryos) if the compound is non-toxic and stable. The FITC label may be degraded in biological fluids. Most applications are in vitro, using the probe to label cells, subcellular structures, or biomolecules for subsequent analysis by fluorescence microscopy, confocal laser scanning microscopy (excitation 495 nm, emission 520 nm), or flow cytometry.

As a fluorescent probe, FITC-Ureidopropionic acid is not typically used in standard enzyme inhibition or receptor binding assays. However, fluorescence intensity detection protocols are well-established for this compound. A representative fluorescence measurement protocol using a fluorescence spectrophotometer is as follows: Prepare FITC-Ureidopropionic acid solutions at known concentrations in an appropriate buffer (e.g., PBS, pH 7.4). Set the excitation wavelength to approximately 495 nm and scan the emission spectrum from 500-600 nm to identify the maximum emission wavelength (typically ~520 nm). Then, measure fluorescence intensity at the optimized excitation/emission pair and construct a standard curve of fluorescence intensity versus concentration. Unknown samples can be quantified by interpolation from this standard curve .

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Cell-free labeling assays: Dissolve FITC-Ureidopropionic acid in DMSO or aqueous buffer (e.g., PBS, pH 7.4) at 1-10 mg/mL. Add to purified proteins (e.g., bovine serum albumin) at a 10- to 50-fold molar excess of the probe to protein. Incubate in the dark at room temperature for 1-4 hours. Remove unbound probe by dialysis, size-exclusion chromatography, or spin filtration. Measure labeling efficiency by absorbance at 495 nm (FITC) and 280 nm (protein) and calculate the degree of labeling (DOL). For direct fluorescence measurement, excite at 495 nm and measure emission at 520 nm using a fluorometer or plate reader. For use as a tracer, prepare working solutions at 1-100 uM in PBS.

FITC-Ureidopropionic acid is widely used in cell-based fluorescence assays for tracking and imaging applications. A standard cell labeling protocol: Culture target cells (e.g., hepatocytes or cancer cell lines) in appropriate medium to desired confluence. Incubate cells with FITC-Ureidopropionic acid (concentration optimization required, typically starting at 1-10 μM) in culture medium for 30-60 minutes at 37°C in a 5% CO₂ incubator. Wash cells thoroughly with PBS to remove unbound probe. Fluorescence signals can then be analyzed using fluorescence microscopy for subcellular localization studies, flow cytometry for quantitative analysis of cellular uptake, or microplate fluorescence readers for high-throughput quantification . For cell tracking experiments, the labeled cells can be monitored over time to assess probe retention and distribution .

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Culture appropriate cell lines (e.g., HeLa, HEK293, or primary cells) in DMEM with 10% FBS. For cell labeling, remove medium, wash cells with PBS, and add FITC-Ureidopropionic acid diluted in serum-free medium or PBS at concentrations of 1-100 uM. Incubate at 37degC for 15-60 minutes in the dark. Wash cells three times with PBS to remove unbound probe. Fix cells with 4% paraformaldehyde for 10 minutes at room temperature (optional). For live-cell imaging, maintain cells in CO2-independent medium or in a heated stage. Visualize using fluorescence microscopy or confocal laser scanning microscopy with FITC filter set (excitation 450-490 nm, emission 515-530 nm). For flow cytometry, detach cells using trypsin-EDTA after washing, resuspend in PBS with 2% FBS, and analyze using a flow cytometer equipped with a 488 nm laser and FL1 (525 nm) detector. Quantify mean fluorescence intensity (MFI) from 10,000 cells per sample. For co-localization studies, co-stain with organelle-specific dyes (e.g., MitoTracker for mitochondria, LysoTracker for lysosomes) and image by confocal microscopy. For kinetic uptake studies, treat cells with FITC-Ureidopropionic acid (10 uM) for various time points (1, 5, 10, 15, 30, 60 minutes), wash, and analyze by flow cytometry to determine uptake rate. For competition assays, pre-incubate cells with excess unlabeled ureidopropionic acid (100-1000 uM) for 15 minutes before adding the fluorescent probe to assess specificity of uptake mechanisms. For metabolism studies, after cell labeling, lyse cells with RIPA buffer, extract, and analyze by LC-MS/MS to confirm the identity of the fluorescent species. Perform appropriate controls including FITC alone and unlabeled cells. Assess cell viability after labeling by propidium iodide staining or MTT to ensure probe concentration is non-toxic.

In vivo animal studies using FITC-Ureidopropionic acid are primarily for biodistribution and metabolic tracking purposes. While specific published protocols are limited, a general approach is described: Administer FITC-Ureidopropionic acid to research animals (e.g., mice or rats) via appropriate routes (intravenous, intraperitoneal, or oral depending on study objectives). At predetermined time points post-administration, collect tissues of interest (liver, kidney, blood, brain, etc.) and prepare homogenates or frozen sections. Fluorescence signals can be visualized using in vivo imaging systems (IVIS) for whole-body distribution, or ex vivo via fluorescence microscopy of tissue sections. Quantitative analysis of probe concentration in tissues can be performed by extracting the fluorescent compound and measuring fluorescence intensity against a standard curve .

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For in vivo imaging in transparent model organisms, dissolve FITC-Ureidopropionic acid in sterile PBS (1-10 mg/mL). For zebrafish embryos (72 hpf), inject 1-10 nL of probe solution into the yolk sac or circulation using a microinjector. Alternatively, add 10-100 uM probe to the embryo water for immersion (if the compound is water-soluble and cell-permeable). Incubate for 30-120 minutes in the dark at 28degC. Wash embryos three times with embryo medium. Anesthetize embryos with tricaine (0.016%) and mount on a glass slide with 1.5% low-melting agarose. Image using a fluorescence stereomicroscope or confocal microscope with FITC filter set. For ex vivo tissue imaging, administer FITC-Ureidopropionic acid to mice via intravenous or intraperitoneal injection (1-10 mg/kg). After 0.5-4 hours, euthanize the animal, harvest organs (liver, kidney, spleen, lung, brain), and prepare frozen sections (10-20 um). Fix sections with 4% paraformaldehyde, mount with DAPI-containing mounting medium, and image by fluorescence microscopy. For tissue distribution studies, homogenize organs, extract, and quantify fluorescence intensity using a plate reader (ex/em 485/525 nm) or normalize to protein concentration. Include vehicle controls and untreated animals to account for autofluorescence. For biodistribution and clearance studies, collect blood at multiple time points (0.5, 1, 2, 4, 8, 24 hours post-injection). Centrifuge to separate plasma, dilute, and measure fluorescence. Use LC-MS/MS to confirm that the fluorescent signal corresponds to intact FITC-Ureidopropionic acid rather than free FITC. Perform toxicity assessment by monitoring body weight, behavior, and histopathology of major organs after 24 hours and 7 days post-injection at the imaging dose. Follow institutional animal care and use committee (IACUC) guidelines for all animal procedures.

Specific pharmacokinetic data for FITC-Ureidopropionic acid (such as half-life, volume of distribution, clearance, bioavailability) is not available in standard literature. As a research-use fluorescent probe, detailed PK profiling is typically not performed for this type of compound. However, the physicochemical properties of the probe can be inferred from its solubility characteristics: the compound is soluble in DMSO and exhibits good water solubility, which may facilitate absorption and distribution . The molecular weight is 563.58 g/mol . Stability data indicates that the probe requires protection from light during storage and handling; powder form is stable at -20°C for up to 3 years, and solutions are stable for 6 months at -80°C or 1 month at -20°C . These properties suggest that in vivo applications would require protection from photobleaching and consideration of the probe's metabolic stability.

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Pharmacokinetic data are not well-established for FITC-Ureidopropionic acid. As a fluorescent probe for in vitro labeling, it is not typically administered systemically. If used in vivo, the FITC label may be cleaved by esterases or other enzymes, potentially releasing free FITC, which has different biodistribution and clearance properties. Free FITC has a plasma half-life of approximately 1-4 hours and is eliminated via renal and biliary excretion. The parent compound, ureidopropionic acid (MW 118.09), is an endogenous metabolite. The probe (MW 563.58, formula C27H25N5O5S) has improved solubility with the FITC conjugate. For in vitro use, dissolve in DMSO (1-10 mM stock) and dilute in water or PBS (final DMSO <0.5%) to working concentrations. Solubility: soluble in DMSO; may dissolve in water at lower concentrations. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month). Protect from light (light-sensitive). Avoid freeze-thaw cycles. For long-term storage of labeled products, store aliquots at -80degC with cryoprotectants. The probe should be handled with care to avoid photobleaching; perform labeling and imaging in subdued light.

According to available product information, FITC-Ureidopropionic acid is for research use only and not for human diagnostic or therapeutic purposes . Specific toxicological data (e.g., LD50, cytotoxicity IC50) is not provided in standard product specifications. General handling precautions include protection from light during transportation and storage, as light exposure may degrade the fluorescent compound . The compound is typically stored as a light yellow to yellow solid powder at -20°C for long-term stability . Users should follow standard laboratory safety practices, including wearing appropriate personal protective equipment, avoiding inhalation and skin contact, and disposing of waste according to institutional regulations. As a fluorescein derivative, the compound generally exhibits low intrinsic toxicity, but individual researchers should conduct appropriate cytotoxicity assays for their specific cell types and experimental conditions.

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The toxicity profile of FITC-Ureidopropionic acid is not well-characterized. As a fluorescent probe, it is intended for in vitro use at low micromolar concentrations (1-100 uM) where it is generally non-toxic to cultured cells as assessed by MTT or propidium iodide viability assays. For in vivo use, limited data exist; FITC-labeled compounds can sometimes cause mild irritation or allergic reactions. The TFA salt is not present in this compound; note that the name "FITC-Ureidopropionic acid" does not include TFA. Standard laboratory precautions (gloves, lab coat, safety glasses) should be used. Avoid inhalation, ingestion, and skin contact. The compound is light-sensitive; store in amber vials. Dispose of waste according to institutional guidelines for fluorescent dyes. In case of accidental eye contact, rinse with water for 15 minutes. For skin contact, wash with soap and water. No genotoxicity or carcinogenicity data are available. This compound is for research use only and not for diagnostic or therapeutic applications.

FITC-Ureidopropionic acid is a fluorescent probe for labeling biomolecules or cells. The FITC (fluorescein isothiocyanate) group is covalently attached to ureidopropionic acid (3-ureidopropionic acid, also known as 3-ureidopropanoic acid or N-carbamoyl-beta-alanine). Ureidopropionic acid is an intermediate in the metabolism of uracil and is also involved in the degradation of pyrimidines and certain amino acids. Under normal physiological conditions, ureidopropionic acid is present in urine and is an endogenous metabolite. The FITC label provides green fluorescence (excitation maximum approximately 495 nm, emission maximum approximately 520 nm), making the probe suitable for fluorescence microscopy, confocal imaging, and flow cytometry. The probe can be used to study the cellular uptake and distribution of ureidopropionic acid or to label proteins and other biomolecules via the carboxylic acid group (amide coupling using EDC/NHS chemistry) or via the FITC isothiocyanate group (which reacts with primary amines to form thiourea bonds). The compound is for research use only and not for diagnostic or therapeutic applications.

C27H25N5O7S

563.58

Ureidopropionic acid;462-88-4

Light yellow to yellow solid powder

FITC-3-Ureidopropionate; FITC-Ureidopropionic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)