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| Targets |
Lipid A component of lipopolysaccharides (LPS). BODIPY TR Cadaverine specifically recognizes and binds to lipid A, the bioactive endotoxin component of LPS on the surface of gram-negative bacteria. The cadaverine moiety mediates binding to the negatively charged lipid A through electrostatic interactions and hydrogen bonding. The BODIPY TR fluorophore provides a red fluorescence signal for detection. This probe does not target a host receptor but rather a bacterial structural component, enabling the detection and study of endotoxin in biological samples. It is used to screen for compounds that can block LPS binding, potentially serving as anti-endotoxin agents.
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| ln Vitro |
The BODIPY TR Cadaverine Assay [1] was used to quantify autophagy in cells. The procedure involved incubating cells for 10 minutes at 125 nM BODIPY TR Cadaverine, followed by four PBS washes and an additional incubation in 10 mM Tris-Cl pH 8 with 0.1% Triton Lysis in X-100. A Molecular Devices SPECTRAmax fluorometer (λex=588nm, λem=616nm) was used to measure the levels of fluorescence. By adding 0.2 mM ethidium bromide and measuring DNA fluorescence (λex=530 nm, λem=590 nm), fluorescence levels were standardized to cell number [1].
In vitro, BODIPY TR Cadaverine is used in fluorescence displacement assays to evaluate the binding affinity of potential LPS-binding agents. At a concentration of 125 nM, the probe binds strongly to LPS, with a dissociation constant (Kd) in the low nanomolar range. The binding is specific for lipid A, and the probe can be competitively displaced by compounds that also bind to lipid A, with a decrease in fluorescence intensity correlating with the displacer's affinity. This assay format is highly sensitive and robust, making it suitable for high-throughput screening of anti-endotoxin agents. BODIPY TR Cadaverine is also used to label LPS for cellular uptake and intracellular trafficking studies, allowing visualization of bacterial endotoxin interactions with immune cells. |
| ln Vivo |
In vivo applications of BODIPY TR Cadaverine are limited, but the probe can be used to label LPS for tracking the biodistribution and clearance of endotoxin in animal models. For example, fluorescently labeled LPS administered intravenously can be visualized in tissues such as the liver and spleen, where it is taken up by Kupffer cells and other phagocytes. The probe may also be used in ex vivo assays with tissue homogenates or serum to detect LPS or screen for anti-endotoxin agents. However, due to its positive charge and propensity to bind negatively charged structures, careful interpretation of in vivo data is required. Detailed in vivo activity data is limited, as the compound is primarily used in vitro and ex vivo.
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| Enzyme Assay |
For a cell-free fluorescence displacement assay, BODIPY TR Cadaverine is first dissolved in DMSO or a suitable buffer (e.g., 10 mM Tris-Cl, pH 7.4) to a stock concentration of 0.1-1 mM. In a 96-well plate, the probe is diluted to a final concentration of 0.1-1 uM in assay buffer (e.g., PBS or Tris-Cl). LPS from E. coli (or other gram-negative bacteria) is added to each well at concentrations ranging from 0.01-100 ug/mL, and the mixture is incubated for 30-60 minutes at room temperature to allow probe binding to LPS. The baseline fluorescence (excitation 580-590 nm, emission 615-625 nm) is measured using a fluorescence plate reader. For the displacement assay, test compounds (potential LPS-binding agents) are added to the probe-LPS mixture at various concentrations (0.01-100 uM), and the decrease in fluorescence intensity (due to displacement of the probe) is measured after an additional 30-60 minutes of incubation. The IC50 (or EC50) for displacement can be calculated by curve fitting.
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| Cell Assay |
A typical cellular protocol for BODIPY TR Cadaverine involves staining cells to quantify autophagy, as the probe accumulates in autophagic vacuoles. Cells (e.g., cardiomyocytes or macrophages) are seeded in 6-well or 24-well plates at 70-80% confluence. BODIPY TR Cadaverine is diluted in culture medium to a final concentration of 125 nM. The cells are incubated with the dye for 10 minutes at 37degC. After incubation, the cells are washed 4 times with PBS to remove unbound dye. To quantify fluorescence, cells are lysed in 500 uL of 10 mM Tris-Cl pH 8.0 containing 0.1% Triton X-100. The lysate is collected, and fluorescence is measured using a fluorimeter or plate reader (excitation 588 nm, emission 616 nm). To normalize for cell number, 0.2 mM ethidium bromide can be added to the lysate, and DNA fluorescence is read at excitation 530 nm and emission 590 nm. Alternatively, protein concentration can be determined by BCA assay. For fluorescence imaging, cells are grown on coverslips, stained as described, washed, and mounted with an anti-fade medium, then visualized using a fluorescence microscope equipped with a Texas Red filter (excitation 560-590 nm, emission 600-650 nm).
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| Animal Protocol |
For in vivo experiments using BODIPY TR Cadaverine-labeled LPS, the probe is first used to label LPS ex vivo. LPS (e.g., 1 mg/mL in PBS) is mixed with BODIPY TR Cadaverine (5-20 uM) and incubated for 1 hour at 37degC. Unbound dye is removed by dialysis or buffer exchange using a centrifugal filter (MWCO 10-30 kDa). The labeled LPS is then diluted in sterile PBS to a final concentration of 0.1-1 mg/mL. Mice (8-12 weeks old, C57BL/6) receive an intravenous injection (tail vein) of 100-200 uL of labeled LPS (dose of 1-5 mg/kg). For kinetics studies, blood is collected via retro-orbital puncture at 0, 5, 15, 30, 60, and 120 minutes post-injection, and plasma fluorescence is measured (excitation 580-590 nm, emission 615-625 nm). At specific time points (1, 4, 8, 24 hours), mice are euthanized, and organs (liver, spleen, kidney, lung, heart) are harvested for fluorescence imaging and quantification. Tissue sections can be prepared for histological analysis to visualize cellular uptake of LPS.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of BODIPY TR Cadaverine are not extensively documented. However, based on its structure and the behavior of similar BODIPY dyes, the compound is expected to be highly lipophilic (estimated logP ~4-6). After intravenous administration, free probe likely binds rapidly to serum proteins (e.g., albumin) and partitions into cell membranes. The plasma half-life is likely short (5-30 minutes) due to rapid clearance by the liver and spleen. When used to label LPS, the PK profile of the labeled LPS will reflect that of native LPS, which is rapidly cleared from circulation by the mononuclear phagocyte system, with a distribution half-life of 2-5 minutes and an elimination half-life of 30-60 minutes. The liver (Kupffer cells) and spleen (macrophages) are the primary sites of accumulation. The labeled LPS is gradually degraded within lysosomes, and the released dye may persist in tissues for longer periods. Detailed quantitative PK data are limited.
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| Toxicity/Toxicokinetics |
Toxicological data for BODIPY TR Cadaverine indicates that it is for research use only and is not intended for human therapeutic use. The compound may be an irritant to the eyes, skin, and respiratory tract. Standard safety precautions, including the use of gloves, a lab coat, and eye protection, should be taken. Avoid inhalation of dust or contact with skin. In vitro cell culture studies using BODIPY TR Cadaverine at a concentration of 125 nM for 10 minutes did not report significant cytotoxicity in cardiomyocytes or other cell types. However, at higher concentrations or longer exposure times, the compound may cause membrane perturbation or phototoxicity. The compound should be stored away from light to minimize photobleaching and potential generation of reactive oxygen species. No chronic toxicity or carcinogenicity data are available.
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| References |
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| Additional Infomation |
BODIPY TR Cadaverine is a research-grade fluorescent probe with no approved clinical applications. It is supplied as a solid (powder) with a purity of ≥95% (typically >98%). The compound is soluble in DMSO, DMF, and ethanol but has limited solubility in aqueous buffers. It should be stored at -20degC in a light-protected container for up to 2-3 years. Stock solutions in DMSO can be stored at -20degC for up to 6 months, but repeated freeze-thaw cycles should be avoided. The probe is widely used in fluorescence displacement assays to identify and characterize LPS-binding agents, as well as to quantify autophagy (by virtue of its accumulation in autophagic vacuoles). The BODIPY TR fluorophore offers high brightness and photostability, making it ideal for long-term imaging studies. Synonyms include BODIPY TR cadaverine, BODIPY TMR cadaverine, and possibly a Texas Red-like conjugate.
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| Molecular Formula |
C26H27BN4O2F2S.HCL
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| Molecular Weight |
544.852
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| Exact Mass |
544.168
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| CAS # |
217190-24-4
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| PubChem CID |
129009841
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| Appearance |
Dark purple to black solid powder
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| Hydrogen Bond Donor Count |
3
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
37
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| Complexity |
906
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| Defined Atom Stereocenter Count |
0
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| SMILES |
Cl.[F-][B+3]1(N2=C(C3SC=CC=3)C=CC2=CC2=CC=C(C3C=CC(OCC(=O)NCCCCCN)=CC=3)[N-]12)[F-]
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| InChi Key |
BKKUIOILPDASKH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H27BF2N4O2S.ClH/c28-27(29)32-20(17-21-9-13-24(33(21)27)25-5-4-16-36-25)8-12-23(32)19-6-10-22(11-7-19)35-18-26(34)31-15-3-1-2-14-30;/h4-13,16-17H,1-3,14-15,18,30H2,(H,31,34);1H
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| Chemical Name |
N-(5-aminopentyl)-2-[4-(2,2-difluoro-12-thiophen-2-yl-1-aza-3-azonia-2-boranuidatricyclo[7.3.0.03,7]dodeca-3,5,7,9,11-pentaen-4-yl)phenoxy]acetamide;hydrochloride
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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 : ~125 mg/mL (~229.42 mM)
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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 | 1.8354 mL | 9.1768 mL | 18.3537 mL | |
| 5 mM | 0.3671 mL | 1.8354 mL | 3.6707 mL | |
| 10 mM | 0.1835 mL | 0.9177 mL | 1.8354 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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