| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
JF525 SE does not target a specific biological receptor. The NHS ester reacts specifically with primary amines under mild conditions (pH 7-9) to form stable amide bonds. The dye itself is a fluorophore for visualization. The SE group is highly reactive toward amines; therefore, the compound is used as a labeling reagent rather than as a ligand.
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| ln Vitro |
In cell-free systems, JF525 SE is characterized by its photophysical properties: absorption max ~525 nm, emission max ~545 nm, high extinction coefficient, and excellent photostability. It is suitable for single-molecule localization microscopy (SMLM) and STED. The NHS ester hydrolyzes in aqueous buffer; the rate of hydrolysis is pH-dependent, with faster hydrolysis at higher pH. Labeling efficiency is measured by fluorescence gel scanning.
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| Enzyme Assay |
A typical protein labeling protocol: Protein (1-10 mg/mL) in PBS (pH 7.2-7.5) is mixed with 5-20 molar excess of JF525 SE (from a DMSO stock). The reaction is incubated at room temperature for 1 hour or at 4degC overnight, protected from light. Unreacted NHS ester is quenched with 50 mM Tris or glycine (pH 7.5). The labeled protein is purified by gel filtration or dialysis. Dye-to-protein ratio is determined by absorbance at 525 nm.
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| Cell Assay |
For cellular labeling, fixed cells are incubated with JF525 SE (1-10 uM) in PBS for 30-60 minutes at room temperature. The dye reacts with surface and intracellular amines. After washing, cells are imaged by fluorescence microscopy. Live-cell labeling is generally not recommended because the NHS ester reacts non-specifically with serum proteins and membrane proteins, causing toxicity and background. Alternatively, the dye can be conjugated to a specific antibody before use.
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| Animal Protocol |
JF525 SE can be conjugated to a targeting antibody, then the conjugate is administered intravenously in mice (e.g., 0.5-2 mg/kg). In vivo imaging is performed using a fluorescence imaging system with excitation/emission filters for green fluorescence. The conjugate's biodistribution is monitored over time (0-48 hours). Alternatively, the free NHS ester is not used in vivo due to rapid hydrolysis and non-specific reactivity. Ex vivo organ imaging confirms target-specific accumulation.
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| ADME/Pharmacokinetics |
As an NHS ester, JF525 SE is unstable in aqueous media; it must be stored desiccated at -20degC. Once conjugated, the labeled protein exhibits PK properties governed by the protein carrier. The free dye has a MW ~700 Da and is rapidly cleared. The dye's high photostability allows long-term imaging. The conjugate should be used within a few hours after preparation or stored frozen at -80degC. The dye is compatible with standard fixation and permeabilization protocols.
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| Toxicity/Toxicokinetics |
JF525 SE is non-toxic at labeling concentrations (≤10 uM for fixed cells). The NHS ester can cause mild irritation if ingested or in contact with skin. In animal studies using conjugates, no significant toxicity has been reported. The compound is for research use only; no clinical toxicity data are available. Standard safety precautions (gloves, lab coat, fume hood) should be followed to avoid exposure.
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| References | |
| Additional Infomation |
JF525 SE is a research-grade fluorescent dye from the Janelia Fluor® series, optimized for high brightness and photostability. It is particularly useful for super-resolution microscopy (STED, PALM). The succinimidyl ester derivative enables efficient and specific labeling of primary amines in proteins and peptides. The dye is not approved for clinical or diagnostic use. It should be stored at -20degC under argon, protected from light, and allowed to warm to room temperature before opening.
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| Molecular Formula |
C31H21F4N3O7
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|---|---|
| Molecular Weight |
623.51
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| Appearance |
Typically exists as solid 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). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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) |
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
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| Solubility (In Vivo) |
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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6038 mL | 8.0191 mL | 16.0382 mL | |
| 5 mM | 0.3208 mL | 1.6038 mL | 3.2076 mL | |
| 10 mM | 0.1604 mL | 0.8019 mL | 1.6038 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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.
Products are for research use only; We do not sell to patients
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