| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
| Enzyme Assay |
Real-time PCR optimization assay using 5(6)-ROX as a fluorescent reporter: Molecular beacons were designed to target a specific DNA sequence, with 5(6)-ROX conjugated as the fluorescent donor at the 5' end and a quencher at the 3' end. The PCR reaction system was prepared by mixing template DNA, primers, molecular beacons, dNTPs, DNA polymerase, buffer, and serial concentrations of 5(6)-ROX (0.1 μM, 0.25 μM, 0.5 μM, 1.0 μM, 2.0 μM, 5.0 μM). The reaction was performed in a real-time PCR instrument with the following conditions: initial denaturation at 95°C for 5 minutes, followed by 40 cycles of denaturation at 95°C for 30 seconds, annealing at 58-62°C (optimized for the target sequence) for 45 seconds, and extension at 72°C for 30 seconds. Fluorescence signals (excitation wavelength ~585 nm, emission wavelength ~605 nm) were collected at the end of each annealing step. The threshold cycle (Ct) values, fluorescence intensity, and specificity (absence of non-specific amplification peaks) were analyzed to determine the optimal concentration of 5(6)-ROX. [1]
|
|---|---|
| References | |
| Additional Infomation |
5(6)-ROX (5(6)-carboxy-X-rhodamine) is a mixture of 5-ROX and 6-ROX, and is a commonly used fluorescent dye. It is often used as a reporter molecule in molecular biology techniques (such as real-time PCR and nucleic acid hybridization) rather than as a therapeutic drug. [1] The literature mainly focuses on optimizing the concentration of 5(6)-ROX in real-time PCR using molecular beacon chemistry to improve the sensitivity and specificity of detection. [1] The main experimental results are: low concentration of 5(6)-ROX (<0.25 μM) leads to weak fluorescence signal and high Ct value (reduced sensitivity); high concentration (>2.0 μM) leads to fluorescence quenching, increased background noise, and may interfere with PCR amplification efficiency. The optimal concentration range is determined to be 0.5-1.0 μM, within which strong specific fluorescence signal, low Ct value and minimal non-specific amplification can be generated. [1] 5(6)-ROX exhibits stable fluorescence under PCR reaction conditions (thermal cycling), has good compatibility with DNA polymerase and other PCR reagents, and has minimal spectral overlap with other commonly used fluorescent dyes (e.g., FAM, TAMRA), making it suitable for multiplex real-time PCR detection. [1]
|
| Molecular Formula |
2[C33H30N2O5]
|
|---|---|
| Molecular Weight |
1069.2034
|
| Exact Mass |
534.215
|
| CAS # |
198978-94-8
|
| PubChem CID |
71312246
|
| Appearance |
Brown to breen solid powder
|
| LogP |
10.96
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
40
|
| Complexity |
1010
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
DAKRRPZULHQSKP-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C33H30N2O5/c36-31(37)20-9-10-21-24(17-20)33(40-32(21)38)25-15-18-5-1-11-34-13-3-7-22(27(18)34)29(25)39-30-23-8-4-14-35-12-2-6-19(28(23)35)16-26(30)33/h9-10,15-17H,1-8,11-14H2,(H,36,37)
|
| Chemical Name |
1-oxospiro[2-benzofuran-3,16'-3-oxa-9,23-diazaheptacyclo[17.7.1.15,9.02,17.04,15.023,27.013,28]octacosa-1(27),2(17),4(15),5(28),13,18-hexaene]-5-carboxylic acid
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~93.53 mM)
|
|---|---|
| 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 | 0.9353 mL | 4.6764 mL | 9.3528 mL | |
| 5 mM | 0.1871 mL | 0.9353 mL | 1.8706 mL | |
| 10 mM | 0.0935 mL | 0.4676 mL | 0.9353 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
Copyright 2020 InvivoChem LLC | All Rights Reserved
