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| Targets |
HBC targets RNA by binding to the Pepper RNA aptamer. The Pepper aptamer is an RNA sequence that specifically recognizes and binds HBC with high affinity. HBC is nonfluorescent in solution, but upon binding to the Pepper aptamer, it undergoes a conformational change that activates its fluorescence. This enables the specific labeling and imaging of RNA molecules in live cells. The high affinity binding (Kd of ~3.5 nM) allows for sensitive detection of RNA targets.
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| ln Vitro |
HBC doesn't glow in solution, but when intramolecular mobility is constrained, it does [1].
In vitro, HBC demonstrates high affinity binding to the Pepper RNA aptamer with a Kd of ~3.5 nM. It is nonfluorescent in solution but emits bright fluorescence upon binding to the Pepper aptamer. The Pepper-HBC 530 complex displays excitation/emission maxima of 485/530 nm. These in vitro characteristics make HBC a valuable tool for RNA imaging in live cells. |
| ln Vivo |
In vivo data for HBC is derived from its use in live cell imaging applications. HBC has been used in the live cell imaging of RNA. When combined with Pepper RNA aptamer, HBC forms a tight complex and activates bright fluorescence, enabling visualization of RNA localization and dynamics in living cells. These applications support its utility as a fluorescent probe for studying RNA biology in live cell models. However, specific published in vivo efficacy studies in animal models are not applicable for this imaging probe.
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| Enzyme Assay |
The in vitro RNA aptamer binding assay for HBC uses the Pepper RNA aptamer. Binding affinity is measured using fluorescence titration, where increasing concentrations of HBC are added to the Pepper aptamer and the increase in fluorescence is monitored. Kd values are calculated from binding curves. The excitation and emission spectra of the Pepper-HBC complex are measured using a fluorescence spectrophotometer.
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| Cell Assay |
Cellular assays for HBC are conducted in live cells expressing the Pepper RNA aptamer. Cells are incubated with HBC, and RNA localization is visualized using fluorescence microscopy or confocal imaging. The Pepper-HBC 530 complex displays excitation/emission maxima of 485/530 nm. The compound's low fluorescence background in the absence of the aptamer allows for specific detection of RNA targets.
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| Animal Protocol |
In vivo imaging studies for HBC are conducted in live cell models and potentially in small animal models expressing the Pepper RNA aptamer. The compound is administered to cells or animals, and fluorescence imaging is performed to visualize RNA localization and dynamics. The Pepper-HBC complex enables real-time monitoring of RNA in living systems. However, specific published in vivo protocols in animal models are not detailed in the available literature. HBC is primarily used as a research tool for RNA imaging.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for HBC is not applicable for this imaging probe as it is used for in vitro and live cell imaging applications rather than systemic drug administration. The compound has a molecular weight of 303.36 g/mol and a molecular formula of C19H17N3O. Storage conditions: store at -20°C. As a fluorescent dye, it is typically used at low concentrations for imaging applications. Detailed PK parameters are not relevant for this research tool.
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| Toxicity/Toxicokinetics |
Toxicity data for HBC is limited as it is used as an imaging probe rather than a therapeutic compound. As with all research compounds, HBC is intended for research use only and not for human therapeutic applications. Standard in vitro cytotoxicity assays would be required to assess potential toxicity in live cell imaging applications. The compound's low fluorescence background makes it suitable for sensitive detection.
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| References | |
| Additional Infomation |
HBC (HBC 530) is a synthetic dye that mimics GFP fluorophore for imaging RNA in live cells. It features a structurally rigid electron acceptor and a strong electron donor. HBC is nonfluorescent in solution but emits bright fluorescence upon binding to Pepper RNA aptamer (Kd of ~3.5 nM). The Pepper-HBC 530 complex displays excitation/emission maxima of 485/530 nm. It has a molecular formula of C19H17N3O and a molecular weight of 303.36 g/mol. HBC is a valuable tool for studying RNA biology in live cells.
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| Molecular Formula |
C19H17N3O
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| Molecular Weight |
303.357784032822
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| Exact Mass |
303.14
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| CAS # |
156840-13-0
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| PubChem CID |
145712177
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| Appearance |
Orange to red solid powder
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| LogP |
2.9
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
23
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| Complexity |
493
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| Defined Atom Stereocenter Count |
0
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| SMILES |
OCCN(C)C1C=CC(/C=C(/C#N)\C2C=CC(C#N)=CC=2)=CC=1
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| InChi Key |
DIAVZHWDYXQAFC-PDGQHHTCSA-N
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| InChi Code |
InChI=1S/C19H17N3O/c1-22(10-11-23)19-8-4-15(5-9-19)12-18(14-21)17-6-2-16(13-20)3-7-17/h2-9,12,23H,10-11H2,1H3/b18-12-
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| Chemical Name |
4-[(E)-1-cyano-2-[4-[2-hydroxyethyl(methyl)amino]phenyl]ethenyl]benzonitrile
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| Synonyms |
HBC 530; HBC
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : 61~125 mg/mL (201.1~412.1 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 | 3.2964 mL | 16.4821 mL | 32.9641 mL | |
| 5 mM | 0.6593 mL | 3.2964 mL | 6.5928 mL | |
| 10 mM | 0.3296 mL | 1.6482 mL | 3.2964 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.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT06122870 | Not yet recruiting | Biological: CampETEC HBC product Biological: ProMilk 85 |
Healthy Volunteer | Johns Hopkins Bloomberg School of Public Health |
December 2023 | Phase 1 |
| NCT04199819 | Recruiting | Drug: entecavir | Liver Transplant; Complications Occult Hepatitis B |
The University of Hong Kong | January 1, 2022 | Not Applicable |
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