| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
Free thiol groups (cysteine residues) on proteins, peptides, and other biomolecules. BODIPY FL L-Cystine contains a disulfide bond (the L-cystine moiety) that can undergo disulfide exchange with free thiols on target molecules. The dye reacts with thiol groups through a disulfide exchange reaction, forming a mixed disulfide between the probe and the target thiol, thereby covalently labeling the target molecule. The reaction is specific for reduced thiols (free -SH) and does not react with oxidized disulfides or other functional groups under mild conditions. This mechanism allows the probe to label proteins that have exposed cysteine residues, such as those in the active sites of enzymes (e.g., phosphatases, proteases) or proteins that undergo redox regulation. The BODIPY FL fluorophore provides a fluorescent signal for detection and quantification. The probe does not have a specific protein target but rather reacts with any accessible thiol, making it a general thiol-labeling reagent. It can also be used to label thiols on membrane proteins or in hydrophobic environments, as the BODIPY FL moiety is relatively lipophilic. The probe can also be used to detect changes in the cellular redox state, such as the ratio of reduced to oxidized glutathione (GSH/GSSG), as the label can be reduced back to free thiols under reducing conditions.
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| ln Vitro |
In vitro, BODIPY FL L-Cystine is used to label proteins or peptides containing free thiol groups. The labeling reaction is typically carried out in buffers at pH 6.5-8.0 (e.g., PBS, Tris-HCl, or HEPES) at room temperature for 1-2 hours or overnight at 4degC. The dye is dissolved in DMSO or DMF to a stock solution of 1-10 mM and then diluted to 1-100 uM in the labeling reaction. The labeling efficiency is dependent on the accessibility of the thiol group and the concentration of the dye and target. The labeled proteins can be analyzed by SDS-PAGE followed by in-gel fluorescence scanning (excitation 488 nm, emission 530 nm), Western blotting (using anti-BODIPY antibodies if available, but typically direct fluorescence detection is used), or by mass spectrometry for identification of labeled peptides. The dye is also used to label thiol groups in cell lysates to assess the thiol proteome or redox state. In live cells, the dye can be used to label surface-exposed thiols on membrane proteins (non-permeant conditions) or total cellular thiols (after permeabilization). However, the dye's ability to cross cell membranes is limited due to the presence of the charged cystine moiety; it may not be efficiently taken up by live cells without permeabilization. The dye is also used in competitive assays to screen for thiol-reactive compounds or to measure the thiol content of purified proteins or biological samples (e.g., by measuring the fluorescence increase upon labeling). It can be used to label membrane proteins, proteins with hydrophobic binding sites, or hydrophobic ligands due to the lipophilic nature of the BODIPY FL group.
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| ln Vivo |
In vivo applications of BODIPY FL L-Cystine are limited due to its reactivity with thiols in the bloodstream and potential non-specific labeling. However, the probe may be used to label thiol-rich tissues or tumors in animal models, particularly in studies of redox status or to track the delivery of thiol-containing drugs or nanoparticles. For example, the dye could be conjugated to a targeting moiety (e.g., an antibody or peptide) via its thiol-reactive group, and then administered to animals for imaging purposes. Alternatively, the dye can be injected locally (e.g., intratumorally) to label thiol-containing proteins in the tumor microenvironment. Detailed in vivo activity data is not widely published, as the probe is primarily used in vitro or ex vivo. Researchers are advised to conduct their own pilot studies to determine in vivo utility. The dye is not approved for human use.
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| Enzyme Assay |
For a cell-free protein labeling reaction, BODIPY FL L-Cystine is dissolved in DMSO to a concentration of 1-10 mM. A purified protein containing free thiol(s) (e.g., BSA, which has a single free cysteine at Cys34) is dissolved in labeling buffer (e.g., 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA). For reduction of disulfide bonds (if needed), the protein is treated with 1-10 mM DTT or TCEP for 30 minutes at 37degC, then the reducing agent is removed by buffer exchange (e.g., using a desalting column) or by dialysis. The protein solution (0.1-1 mg/mL) is incubated with BODIPY FL L-Cystine (1-20 molar excess over protein thiols) for 1-2 hours at room temperature, protected from light. The reaction is quenched by the addition of excess cysteine or beta-mercaptoethanol (10-20 mM) for 10 minutes. Unreacted dye is removed by buffer exchange (desalting column, dialysis, or precipitation). The labeling efficiency is assessed by measuring the absorbance at 504 nm (for BODIPY) and 280 nm (for protein) using a spectrophotometer, or by comparing the fluorescence intensity of the labeled protein to a standard curve of the free dye. The labeled protein is analyzed by SDS-PAGE and in-gel fluorescence scanning (excitation 488 nm, emission 530 nm) to confirm labeling and assess purity. For determining the thiol content of a sample (e.g., cell lysate, biological fluid), BODIPY FL L-Cystine can be used in a fluorescence-based assay. The sample (containing reduced thiols) is incubated with the dye (10-50 uM) in buffer for 30-60 minutes, and the increase in fluorescence (excitation 504 nm, emission 511-530 nm) is measured. The concentration of thiols is determined by comparison to a standard curve generated using reduced glutathione (GSH) or cysteine standards. The dye may also be used to label thiol groups on the surface of nanoparticles or other materials for bio-conjugation studies.
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| Cell Assay |
For labeling thiol groups on the surface of live cells (non-permeant), cells (e.g., HeLa, CHO) are grown to 70-80% confluence in culture dishes or on coverslips. The culture medium is removed, and cells are washed twice with PBS (pH 7.4) to remove serum proteins (which contain thiols and would consume the dye). A solution of BODIPY FL L-Cystine (1-10 uM) in PBS (without reducing agents) is prepared fresh from a DMSO stock (final DMSO <0.5%). The dye solution is added to the cells, and the cells are incubated for 15-60 minutes at room temperature or 4degC (to minimize endocytosis). After incubation, cells are washed 3 times with PBS to remove unbound dye. For fluorescence imaging, cells can be fixed with 4% paraformaldehyde for 10-15 minutes, washed, and mounted with an anti-fade mounting medium. Images are acquired using a fluorescence microscope with excitation at 488 nm and emission at 505-550 nm (FITC channel). For labeling intracellular thiols, cells must first be permeabilized. Cells are fixed with 4% paraformaldehyde (10-15 min, RT) and then permeabilized with 0.1-0.5% Triton X-100 in PBS for 5-10 minutes. After washing, cells are incubated with the dye as described above. Alternatively, live cells can be treated with a reducing agent (e.g., TCEP) to reduce disulfide bonds and unmask additional thiols, then labeled after washing out the reducing agent. For flow cytometry analysis, cells are stained in suspension as described, washed, and analyzed on a flow cytometer equipped with a 488 nm laser and a 530/30 nm emission filter (FITC channel). The dye can also be used to label thiols in cell lysates or tissue homogenates. For ex vivo labeling of tissue sections, frozen or paraffin-embedded sections are deparaffinized, rehydrated, washed with PBS, and then incubated with the dye (1-10 uM in PBS) for 30-60 minutes at room temperature. After washing, sections are mounted and imaged. The dye should be protected from light at all steps to prevent photobleaching.
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| Animal Protocol |
In vivo labeling protocols for BODIPY FL L-Cystine are not well-established. For potential systemic administration, the dye is dissolved in sterile PBS (containing <5% DMSO) at a concentration of 0.5-2 mg/mL, filtered through a 0.22 um filter, and administered intravenously (tail vein) to mice at a dose of 0.1-1 mg/kg. After circulation for 30-120 minutes, mice are euthanized, and tissues are collected for ex vivo fluorescence imaging (excitation 488 nm, emission 510-530 nm) and histological analysis. However, non-specific binding to serum albumin and other thiol-containing proteins in the bloodstream may limit the utility of systemic administration. Alternative routes include intratumoral or intradermal injection for local labeling. The dye can also be conjugated to a targeting ligand (e.g., antibody, peptide, or aptamer) via a thiol-maleimide coupling or disulfide exchange, and the conjugate is then administered for targeted imaging. For example, an antibody is first reduced (using TCEP or DTT) to generate free thiols, then reacted with BODIPY FL L-Cystine (or a maleimide derivative of BODIPY FL) to produce a labeled antibody. The labeled antibody is purified and administered to mice (e.g., 0.5-5 mg/kg, IV) for in vivo tumor imaging. Fluorescence imaging is performed 4-24 hours post-injection using a whole-body imaging system with appropriate filters (excitation 470-490 nm, emission 510-540 nm). Due to green fluorescence, signal penetration is limited to superficial tissues, and higher background may be observed compared to near-infrared dyes. Researchers should optimize the protocol for their specific application. The compound is for research use only and not for human use.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for BODIPY FL L-Cystine are limited. As a thiol-reactive dye, it is expected to bind rapidly to plasma proteins (especially albumin) and to cellular thiols upon systemic administration. The free probe (unbound) has a molecular weight of 788.45 and is moderately lipophilic (estimated logP ~2-3). After intravenous injection, the compound is rapidly cleared from the circulation with a half-life of minutes, as it reacts with thiols or is taken up by the liver and spleen. When conjugated to a targeting ligand (e.g., antibody), the PK profile will be primarily determined by the ligand, with a longer half-life (hours to days) and different biodistribution. In vitro, the dye is relatively stable in the dark and in the absence of reducing agents, but it can be reduced by DTT or TCEP, which may be relevant for intracellular redox studies. For quantitative analysis, the dye can be extracted from biological samples (e.g., plasma, tissues) by organic solvent (e.g., methanol/chloroform) and analyzed by HPLC with fluorescence detection, using an internal standard. Researchers are advised to perform custom PK studies if required for their specific application, as detailed public data is not available.
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| Toxicity/Toxicokinetics |
Toxicological data for BODIPY FL L-Cystine are limited. In vitro, the dye is generally non-toxic at concentrations used for labeling (1-10 uM) over short exposure times (1-2 hours), as assessed by cell viability assays (MTT, LDH). At higher concentrations (e.g., >50 uM) or prolonged exposure (>24 hours), some cytotoxicity may be observed due to the effects of thiol modification on cellular redox balance or protein function. The dye is for research use only and is not intended for human use. Standard safety precautions for handling organic dyes and chemicals should be followed: use gloves, lab coat, and eye protection; avoid inhalation of dust; work in a well-ventilated area; protect from light. The compound should be stored at -20degC, protected from light and moisture, and should be allowed to warm to room temperature before opening. Proper disposal of the compound and any waste should be in accordance with local regulations. No genotoxicity, carcinogenicity, or reproductive toxicity data are available.
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| References | |
| Additional Infomation |
BODIPY FL L-Cystine is a research-grade thiol-reactive fluorescent dye with no approved clinical applications. It is supplied as a solid powder with a purity of ≥95% (typically >95%). The molecular weight is 788.45, and the formula is C34H38B2F4N6O6S2. The dye is soluble in DMSO (10-20 mg/mL) and other organic solvents (DMF, ethanol), but has limited aqueous solubility. Stock solutions (1-10 mM) in DMSO should be stored at -20degC, protected from light, for up to 6 months. It can be used to label membrane proteins, proteins with hydrophobic binding sites, or hydrophobic ligands. The BODIPY FL fluorophore has excitation/emission maxima at 504/511 nm, producing a bright green fluorescence. The L-cystine moiety provides specificity for thiol groups. The probe is particularly useful for studying protein-ligand interactions, cellular redox status, and cysteine-related processes. It can also be used to label thiol groups on the surface of cells or to label purified proteins for biochemical assays. Synonyms include BODIPY FL L-cystine. For detailed protocols, refer to the product data sheet and primary literature. The compound is intended for research use only and is not for diagnostic or therapeutic use.
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| Molecular Formula |
C34H36B2N6O6F4S2-2.H+
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|---|---|
| Molecular Weight |
787.44
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| Exact Mass |
788.241
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| CAS # |
1180490-57-6
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| PubChem CID |
168008656
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| Appearance |
Orange to red solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
15
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| Heavy Atom Count |
54
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| Complexity |
1670
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C(C1=CC=C2C=C3C(C)=CC(C)=N3[B+3]([F-])([F-])[N-]12)CC(=O)NC(C(=O)[O-])CSSCC(C(=O)[O-])NC(=O)CCC1=CC=C2C=C3C(C)=CC(C)=N3[B+3]([F-])([F-])[N-]12.[H+]
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| InChi Key |
IUOBNSXKYBKQEY-NSOVKSMOSA-N
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| InChi Code |
InChI=1S/C34H38B2F4N6O6S2/c1-19-13-21(3)43-29(19)15-25-7-5-23(45(25)35(43,37)38)9-11-31(47)41-27(33(49)50)17-53-54-18-28(34(51)52)42-32(48)12-10-24-6-8-26-16-30-20(2)14-22(4)44(30)36(39,40)46(24)26/h5-8,13-16,27-28H,9-12,17-18H2,1-4H3,(H,41,47)(H,42,48)(H,49,50)(H,51,52)/t27-,28-/m0/s1
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| Chemical Name |
(2R)-3-[[(2R)-2-carboxy-2-[3-(2,2-difluoro-10,12-dimethyl-1-aza-3-azonia-2-boranuidatricyclo[7.3.0.03,7]dodeca-3,5,7,9,11-pentaen-4-yl)propanoylamino]ethyl]disulfanyl]-2-[3-(2,2-difluoro-10,12-dimethyl-1-aza-3-azonia-2-boranuidatricyclo[7.3.0.03,7]dodeca-3,5,7,9,11-pentaen-4-yl)propanoylamino]propanoic acid
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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: 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 : ~50 mg/mL (~63.42 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.67 mg/mL (2.12 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2699 mL | 6.3497 mL | 12.6994 mL | |
| 5 mM | 0.2540 mL | 1.2699 mL | 2.5399 mL | |
| 10 mM | 0.1270 mL | 0.6350 mL | 1.2699 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.
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