| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
The primary target of 6-ROX is not a biological receptor or enzyme, but rather the oligonucleotide or biomolecule to which it is conjugated for detection purposes. As a fluorescent labeling reagent, 6-ROX is used to label nucleotides and oligonucleotides for DNA sequencing and FRET imaging applications. The carboxy group on the compound can be activated to form reactive esters that react with amines on nucleotides or other biomolecules. The labeled oligonucleotides can then be used as probes in various fluorescence-based assays, including FRET imaging, where 6-ROX acts as an acceptor molecule coupled to 5-FAM as the donor.
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| ln Vitro |
In vitro, 6-ROX is used as a fluorescent marker for oligonucleotides and nucleotides. It is predominantly used to label nucleotides and for DNA sequencing. The compound has excitation/emission wavelengths of 570/591 nm. It acts as an acceptor molecule coupled to 5-FAM as the donor in FRET imaging. The labeled oligonucleotides can be used as probes in various fluorescence-based assays, including DNA sequencing, PCR, and hybridization assays. The compound's fluorescence properties allow for sensitive detection of target nucleic acids in complex biological samples.
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| ln Vivo |
In vivo studies of 6-ROX are limited, as the compound is primarily used as an in vitro research reagent for labeling and detection. However, fluorescent dyes like 6-ROX may be used in animal models for imaging applications, such as tracking oligonucleotides or cells in vivo. In such studies, labeled oligonucleotides are administered to animals, and their distribution and fate are monitored by fluorescence imaging. The compound's fluorescence properties enable sensitive detection in biological samples. No extensive in vivo pharmacological or toxicological studies have been reported.
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| Enzyme Assay |
For fluorescence characterization, 6-ROX is dissolved in an appropriate solvent (e.g., DMSO, methanol, or aqueous buffer) and its absorption and emission spectra are measured using a UV-Vis spectrophotometer and fluorescence spectrometer, respectively. Excitation and emission maxima are determined (Ex=570 nm, Em=591 nm). For labeling reactions, the carboxy group of 6-ROX is activated to an NHS ester using EDC and NHS. The activated ester is then reacted with the target oligonucleotide or nucleotide containing a primary amine group in a buffer at pH 7.0-8.5 for 1-2 hours at room temperature. The labeled product is purified by HPLC or other methods. The labeling efficiency is assessed by measuring the absorbance or fluorescence of the labeled product.
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| Cell Assay |
For cellular studies, 6-ROX-labeled oligonucleotides or probes are used to stain cells or to detect specific nucleic acid sequences in cells. Cells are cultured and fixed, then incubated with the labeled probe. After washing, cells are analyzed by fluorescence microscopy or flow cytometry using appropriate filter sets for excitation at 570 nm and emission at 591 nm. For FRET imaging, 6-ROX-labeled probes are used in combination with 5-FAM-labeled probes to study molecular interactions.
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| Animal Protocol |
For in vivo imaging studies, 6-ROX-labeled oligonucleotides are administered to animals via intravenous or intraperitoneal injection. The distribution and fate of the labeled molecules are monitored using an in vivo imaging system equipped with appropriate excitation and emission filters. Tissues and organs may be collected at various time points for ex vivo fluorescence imaging or homogenization for fluorometric analysis. However, such studies are not typical for this compound.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for 6-ROX are not available, as the compound is used as a research reagent rather than a therapeutic agent. If used in vivo, the compound would be expected to have distribution to various tissues. However, no dedicated ADME studies have been reported. The compound is primarily used in vitro and is not intended for in vivo pharmacokinetic characterization.
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| Toxicity/Toxicokinetics |
Toxicological data for 6-ROX are limited, as the compound is used as a research reagent. The compound is generally considered to have low toxicity at the concentrations used for labeling and staining applications. No acute toxicity, mutagenicity, or carcinogenicity data have been reported. As with all chemical reagents, appropriate safety precautions should be taken when handling 6-ROX, including the use of personal protective equipment and work in a well-ventilated area.
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| Additional Infomation |
6-ROX (CAS 194785-18-7), also known as 6-Carboxy-X-rhodamine, is a fluorescent oligonucleotide marker used as an acceptor in FRET imaging. It has a molecular formula of C₃₃H₃₀N₂O₅ and a molecular weight of 534.60. The compound is used to label nucleotides and for DNA sequencing. It has excitation/emission of 570/591 nm. 6-ROX acts as an acceptor coupled to 5-FAM as the donor in FRET imaging. The compound is a rhodamine-class fluorescent dye. It is strictly for research use only.
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| Molecular Formula |
C33H30N2O5
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|---|---|
| Molecular Weight |
534.601699999999
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| Exact Mass |
534.215
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| CAS # |
194785-18-7
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| Related CAS # |
6-ROX hydrochloride;1689512-96-6
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| PubChem CID |
2762612
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| Appearance |
Brown to black solid powder
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| LogP |
5.123
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
40
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| Complexity |
1200
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
WQZIDRAQTRIQDX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C33H30N2O5/c36-32(37)20-9-10-21(33(38)39)24(17-20)27-25-15-18-5-1-11-34-13-3-7-22(28(18)34)30(25)40-31-23-8-4-14-35-12-2-6-19(29(23)35)16-26(27)31/h9-10,15-17H,1-8,11-14H2,(H-,36,37,38,39)
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| Chemical Name |
4-carboxy-3-(3-oxa-23-aza-9-azoniaheptacyclo[17.7.1.15,9.02,17.04,15.023,27.013,28]octacosa-1(27),2(17),4,9(28),13,15,18-heptaen-16-yl)benzoate
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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 : ~25 mg/mL (~46.76 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (1.87 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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. Solubility in Formulation 2: ≥ 1 mg/mL (1.87 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8706 mL | 9.3528 mL | 18.7056 mL | |
| 5 mM | 0.3741 mL | 1.8706 mL | 3.7411 mL | |
| 10 mM | 0.1871 mL | 0.9353 mL | 1.8706 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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